Mobile website
Volume 20,Issue 3,2013 Table of Contents
2013, 20(3):259-265.
DOI: 10.3872/j.issn.1007-385X.2013.03.001
Abstract:
Increasing evidence indicated that tumor inflammatory microenviroment plays an important role in cancer initiation and development. The signaling pathways mediated by inflammatory cytokines participate in malignant transformation of tumor cells. Epithelial-mesenchymal transition (EMT) is a critical mechanism underlying transformation of tumor cells. A network composed by inflammatory cytokines (interleukin, TNF-α and TGF-β), key transcription factors of EMT, including zinc finger E-box-binding homeobox (ZEB), Snail and Twist, and some miRNAs (let-7 and miR-200) regulates malignant transformation of tumor, generation of drug resistance and the proliferation and the self-renewal of cancer stem cell (CSC). This article reviews the recent studies on the molecular mechanisms, by which inflammation and related signaling pathways regulate tumor initiation, development and CSC formation.
Increasing evidence indicated that tumor inflammatory microenviroment plays an important role in cancer initiation and development. The signaling pathways mediated by inflammatory cytokines participate in malignant transformation of tumor cells. Epithelial-mesenchymal transition (EMT) is a critical mechanism underlying transformation of tumor cells. A network composed by inflammatory cytokines (interleukin, TNF-α and TGF-β), key transcription factors of EMT, including zinc finger E-box-binding homeobox (ZEB), Snail and Twist, and some miRNAs (let-7 and miR-200) regulates malignant transformation of tumor, generation of drug resistance and the proliferation and the self-renewal of cancer stem cell (CSC). This article reviews the recent studies on the molecular mechanisms, by which inflammation and related signaling pathways regulate tumor initiation, development and CSC formation.
2013, 20(3):266-271.
DOI: 10.3872/j.issn.1007-385X.2013.03.002
Abstract:
Objective:To explore the inhibitory effect of mouse phosphatase of regenerating liver-3 (mPRL-3)-targeted DNA vaccine on the growth of mouse breast cancer D2F2 cells in vivo. Methods: The eukaryotic expression vector pVAX1-mPRL-3 targeting mPRL-3 was constructed and transfected into quail fibroblasts QM7 cells. The mPRL-3 protein expression in QM7 cells was detected by Western blotting. D2F2 mouse breast cancer cells were infected with recombinant lentivirus (Lv-mPRL-3) or control vector (Lv-Ctrl) to generate cells expressing mPRL-3 (mPRL-3-D2F2) or control cells (NC-D2F2), and the expression of mPRL-3 protein in mouse breast cancer D2F2 cells was analyzed by Western blotting. The mPRL-3-D2F2 and NC-D2F2 cells were respectively inoculated into BALB/c mice’s left mammary fad pat, then the BALB/c mice were immunized with pVAX1-mPRL-3 DNA vaccine (mPRL-3-D2F2/pVAX1-mPRL-3) by gene gun, and mPRL-3-D2F2/pVAX1-Ctrl and NC-D2F2/pVAX1-mPRL-3 were set as controls. The volume of tumor and the survival time of mice were monitored. Results: The eukaryotic expression vector pVAX1-mPRL-3 was identified by enzymatic digestion and DNA sequencing and the expression of mPRL-3 protein in QM7 cells was also confirmed. Western blotting assay results showed that the over-expression of mPRL-3 protein was detected in mPRL-3-D2F2, but not in NC-D2F2 cells. The tumor volume of the tumor-bearing mice immunized with pVAX1-mPRL-3 vaccine was significantly lower than that of the control group (\[835.3±509.8\] vs \[1543.0±578.4\] mm3, P<001\]. The pVAX1-mPRL-3 vaccination could significantly prolong the survival time of the tumor bearing mice (median survival time: 55.5 vs 38 d, P<005). Conclusion: PRL-3 -targeted vaccination mediated by gene gun can inhibit the growth of mouse breast cancer D2F2 cell xenografts expressing mPRL-3, which could be a potential therapy strategy for PRL-3 positive tumor.
Objective:To explore the inhibitory effect of mouse phosphatase of regenerating liver-3 (mPRL-3)-targeted DNA vaccine on the growth of mouse breast cancer D2F2 cells in vivo. Methods: The eukaryotic expression vector pVAX1-mPRL-3 targeting mPRL-3 was constructed and transfected into quail fibroblasts QM7 cells. The mPRL-3 protein expression in QM7 cells was detected by Western blotting. D2F2 mouse breast cancer cells were infected with recombinant lentivirus (Lv-mPRL-3) or control vector (Lv-Ctrl) to generate cells expressing mPRL-3 (mPRL-3-D2F2) or control cells (NC-D2F2), and the expression of mPRL-3 protein in mouse breast cancer D2F2 cells was analyzed by Western blotting. The mPRL-3-D2F2 and NC-D2F2 cells were respectively inoculated into BALB/c mice’s left mammary fad pat, then the BALB/c mice were immunized with pVAX1-mPRL-3 DNA vaccine (mPRL-3-D2F2/pVAX1-mPRL-3) by gene gun, and mPRL-3-D2F2/pVAX1-Ctrl and NC-D2F2/pVAX1-mPRL-3 were set as controls. The volume of tumor and the survival time of mice were monitored. Results: The eukaryotic expression vector pVAX1-mPRL-3 was identified by enzymatic digestion and DNA sequencing and the expression of mPRL-3 protein in QM7 cells was also confirmed. Western blotting assay results showed that the over-expression of mPRL-3 protein was detected in mPRL-3-D2F2, but not in NC-D2F2 cells. The tumor volume of the tumor-bearing mice immunized with pVAX1-mPRL-3 vaccine was significantly lower than that of the control group (\[835.3±509.8\] vs \[1543.0±578.4\] mm3, P<001\]. The pVAX1-mPRL-3 vaccination could significantly prolong the survival time of the tumor bearing mice (median survival time: 55.5 vs 38 d, P<005). Conclusion: PRL-3 -targeted vaccination mediated by gene gun can inhibit the growth of mouse breast cancer D2F2 cell xenografts expressing mPRL-3, which could be a potential therapy strategy for PRL-3 positive tumor.
2013, 20(3):272-277.
DOI: 10.3872/j.issn.1007-385X.2013.03.003
Abstract:
Objective:To study the radiosensitization effect of homocysteine-inducible, endoplasmic reticulum stress-inducible, ubiquitin-like domain member 1 ( HerpUD1 ) on human liver L-02 cells through silencing the expression of HerpUD1 gene by RNA interference technology. Methods: Based on the HerpUD1 gene sequence, shRNA expression vector interfering HerpUD1 gene was constructed and L-02 cells were stably transfected. The expressions of HerpUD1 mRNA and protein in cells after transfection were detected by real-time PCR and Western blotting. The transfected cells were radiated by 8 Gy X ray and the cell apoptosis was analyzed by Hoechst fluorescence assay. Results: Eukaryotic expression interference vector targeting HerpUD1 gene was successfully constructed and L-02 cell line stably transfected with shHerpUD1 was established. The shHerpUD1-1420 interference fragment had the highest interference efficiency in stably transfected L-02 cells. Compared with pGPU6-NC (empty vector) group, the expression level of HerpUD1 mRNA in shHerpUD1-1420 group was significantly decreased (\[0.15±005\] vs \[1.00±0.08\], P<0.05), and the expression level of HerpUD1 protein was also significantly reduced (\[0.19±0.01\] vs \[1.62±0.08\], P<0.01). The apoptotic rate was higher in L-02-shHerpUD1-1420 group than that in L-02-NC group (negative control group) 24 h after 8 Gy X ray irradiation (\[1023±3.37\]% vs \[6.89±1.49\]%, P<0.05), while there was no significant difference between L-02-NC and L-02-Neo group. Conclusion:Silencing the expression of HerpUD1 gene significantly increases the X ray radiation-induced apoptosis of L-02 cells, therefore playing an role in radiosensitization.
Objective:To study the radiosensitization effect of homocysteine-inducible, endoplasmic reticulum stress-inducible, ubiquitin-like domain member 1 ( HerpUD1 ) on human liver L-02 cells through silencing the expression of HerpUD1 gene by RNA interference technology. Methods: Based on the HerpUD1 gene sequence, shRNA expression vector interfering HerpUD1 gene was constructed and L-02 cells were stably transfected. The expressions of HerpUD1 mRNA and protein in cells after transfection were detected by real-time PCR and Western blotting. The transfected cells were radiated by 8 Gy X ray and the cell apoptosis was analyzed by Hoechst fluorescence assay. Results: Eukaryotic expression interference vector targeting HerpUD1 gene was successfully constructed and L-02 cell line stably transfected with shHerpUD1 was established. The shHerpUD1-1420 interference fragment had the highest interference efficiency in stably transfected L-02 cells. Compared with pGPU6-NC (empty vector) group, the expression level of HerpUD1 mRNA in shHerpUD1-1420 group was significantly decreased (\[0.15±005\] vs \[1.00±0.08\], P<0.05), and the expression level of HerpUD1 protein was also significantly reduced (\[0.19±0.01\] vs \[1.62±0.08\], P<0.01). The apoptotic rate was higher in L-02-shHerpUD1-1420 group than that in L-02-NC group (negative control group) 24 h after 8 Gy X ray irradiation (\[1023±3.37\]% vs \[6.89±1.49\]%, P<0.05), while there was no significant difference between L-02-NC and L-02-Neo group. Conclusion:Silencing the expression of HerpUD1 gene significantly increases the X ray radiation-induced apoptosis of L-02 cells, therefore playing an role in radiosensitization.
2013, 20(3):278-283.
DOI: 10.3872/j.issn.1007-385X.2013.03.004
Abstract:
Objective:To explore the potentiation of silencing atxia-telangiectasia mutated (ATM) gene expression in the radiosensitization effect of cytosine-phophate-guanine oligodeoxynucleotide (CpG ODN) 7909 on non-small cell lung cancer A549 cell line. Methods: ATM-siRNA was transfected into A549 cells, and the expression of ATM protein in A549 cells was examined by Western blotting. A549 cells were randomly classified into six groups: control group, CpG group, X-ray (IR) group, CpG+IR group, ATM-siRNA+CpG+IR group and NC-siRNA+CpG+IR group. Cell colony rates were evaluated by colony formation assay. One-hit multi-target model and linear quadratic model, which generated the radiation dose survival curve of the A549 cells, were fitted with Graphpad prism 5.0 software, and the parameters, including D0, Dq, N, α/β and SF2 were applied to analyze radiation damage repair capacity of A549 cells. The apoptosis of A549 cells was analyzed by flow cytometry. Results: ATM-siRNA transfection remarkably inhibited the expression of ATM protein in A549 cells (P<001). X-ray radiation inhibited the colony formation capacity of A549 cells in a dose-dependent manner (P<0.05). Moreover, a further decrease of the colony formation capacity of A549 cells was found in CpG+IR radiation combined treatment group (P<0.01). With the transfection of ATM-siRNA, the colony formation capacity of CpG-treated A549 cells was further decreased (10 Gy, \[0.05±0.00\]% vs \[0.71±0.00\]%, P<0.01). Radiation damage dose survival curves demonstrated that the value of α/β was significantly increased (1.48 vs 0.97，P<005)and the radiation damage repair capability was significantly decreased in ATM-siRNA+CpG+IR group compared with CpG+IR group. The apoptotic rate was significantly increased in CpG+IR group compared with IR radiation group (\[9.18±016\]% vs \[6.56±0.33\]%, P<0.01). The apoptotic rate of A549 cells in ATM-siRNA+CpG+IR group was further increased (\[9.18±0.16\]% vs \[10.45±0.40\]%, P<0.05). Conclusion:Silencing expression of ATM by siRNA can enhance the radiosensitization effect of CpG ODN 7909 on A549 cells. ATM may serve as a potential target for gene therapy of lung cancer.
Objective:To explore the potentiation of silencing atxia-telangiectasia mutated (ATM) gene expression in the radiosensitization effect of cytosine-phophate-guanine oligodeoxynucleotide (CpG ODN) 7909 on non-small cell lung cancer A549 cell line. Methods: ATM-siRNA was transfected into A549 cells, and the expression of ATM protein in A549 cells was examined by Western blotting. A549 cells were randomly classified into six groups: control group, CpG group, X-ray (IR) group, CpG+IR group, ATM-siRNA+CpG+IR group and NC-siRNA+CpG+IR group. Cell colony rates were evaluated by colony formation assay. One-hit multi-target model and linear quadratic model, which generated the radiation dose survival curve of the A549 cells, were fitted with Graphpad prism 5.0 software, and the parameters, including D0, Dq, N, α/β and SF2 were applied to analyze radiation damage repair capacity of A549 cells. The apoptosis of A549 cells was analyzed by flow cytometry. Results: ATM-siRNA transfection remarkably inhibited the expression of ATM protein in A549 cells (P<001). X-ray radiation inhibited the colony formation capacity of A549 cells in a dose-dependent manner (P<0.05). Moreover, a further decrease of the colony formation capacity of A549 cells was found in CpG+IR radiation combined treatment group (P<0.01). With the transfection of ATM-siRNA, the colony formation capacity of CpG-treated A549 cells was further decreased (10 Gy, \[0.05±0.00\]% vs \[0.71±0.00\]%, P<0.01). Radiation damage dose survival curves demonstrated that the value of α/β was significantly increased (1.48 vs 0.97，P<005)and the radiation damage repair capability was significantly decreased in ATM-siRNA+CpG+IR group compared with CpG+IR group. The apoptotic rate was significantly increased in CpG+IR group compared with IR radiation group (\[9.18±016\]% vs \[6.56±0.33\]%, P<0.01). The apoptotic rate of A549 cells in ATM-siRNA+CpG+IR group was further increased (\[9.18±0.16\]% vs \[10.45±0.40\]%, P<0.05). Conclusion:Silencing expression of ATM by siRNA can enhance the radiosensitization effect of CpG ODN 7909 on A549 cells. ATM may serve as a potential target for gene therapy of lung cancer.
2013, 20(3):284-288.
DOI: 10.3872/j.issn.1007-385X.2013.03.005
Abstract:
Objective:To explore the effect of IL-22 on the proliferation of colorectal cancer cells and to illustrate its underlying molecular mechanism. Methods: The relative expression levels of IL-22 receptor 1 ( IL-22R1 ) mRNA in colorectal cancer SW480 and SW620 cells were detected by real-time PCR. The different mass concentrations of IL-22 (0, 1, 5, 10 ng/ml) were used to treat colorectal cancer cells. The effect of IL-22 on the proliferation of SW480 and SW620 cells was analyzed by MTT assay and colony formation assay. SW480 and SW620 cells were stimulated with IL-22 for various time points and were subjected to Western blotting for the detection of protein phosphorylations, including STAT3, AKT, ERK, JNK and P38. Whether the specific inhibitor LLL12 blocking phosphorylation of STAT3 could promote the proliferation of colorectal cancer cells induced by IL-22 was observed. Results: IL-22R1 mRNA was expressed in colorectal cancer cells (SW480, SW620). IL-22 can promote the proliferation of SW480 and SW620 cells in a dose-dependent manner. After the treatment with IL-22 (10 ng/ml), the cell proliferation folds were significantly increased (SW480: \[5.18±0.212\] vs \[2.64±0.27\], SW620: \[814±0.61\] vs \[6.08±0.096\], P<0.01). Moreover, IL-22 enanced the colony formation ability of SW480 and SW620 cells. The colony numbers of IL-22 treating group were significantly higher compared with the control group (\[1 680.67±124.05\] vs \[730±64.29\], \[2 668±116.37\] vs \[1 294±171.61\], P<0.01). Western blotting demonstrated that STAT3 pathway can be activated after the treatment of IL-22. However, there was no significant alteration in AKT, ERK, JNK, P38 pathways. The proliferative effect of IL-22 can be attenuated after the treatment of STAT3 phosphorylation inhibitor LLL12. Conclusion: IL-22 may promote the proliferation of colon cancer SW480 and SW620 cells via STAT3 signaling pathway.
Objective:To explore the effect of IL-22 on the proliferation of colorectal cancer cells and to illustrate its underlying molecular mechanism. Methods: The relative expression levels of IL-22 receptor 1 ( IL-22R1 ) mRNA in colorectal cancer SW480 and SW620 cells were detected by real-time PCR. The different mass concentrations of IL-22 (0, 1, 5, 10 ng/ml) were used to treat colorectal cancer cells. The effect of IL-22 on the proliferation of SW480 and SW620 cells was analyzed by MTT assay and colony formation assay. SW480 and SW620 cells were stimulated with IL-22 for various time points and were subjected to Western blotting for the detection of protein phosphorylations, including STAT3, AKT, ERK, JNK and P38. Whether the specific inhibitor LLL12 blocking phosphorylation of STAT3 could promote the proliferation of colorectal cancer cells induced by IL-22 was observed. Results: IL-22R1 mRNA was expressed in colorectal cancer cells (SW480, SW620). IL-22 can promote the proliferation of SW480 and SW620 cells in a dose-dependent manner. After the treatment with IL-22 (10 ng/ml), the cell proliferation folds were significantly increased (SW480: \[5.18±0.212\] vs \[2.64±0.27\], SW620: \[814±0.61\] vs \[6.08±0.096\], P<0.01). Moreover, IL-22 enanced the colony formation ability of SW480 and SW620 cells. The colony numbers of IL-22 treating group were significantly higher compared with the control group (\[1 680.67±124.05\] vs \[730±64.29\], \[2 668±116.37\] vs \[1 294±171.61\], P<0.01). Western blotting demonstrated that STAT3 pathway can be activated after the treatment of IL-22. However, there was no significant alteration in AKT, ERK, JNK, P38 pathways. The proliferative effect of IL-22 can be attenuated after the treatment of STAT3 phosphorylation inhibitor LLL12. Conclusion: IL-22 may promote the proliferation of colon cancer SW480 and SW620 cells via STAT3 signaling pathway.
2013, 20(3):289-294.
DOI: 10.3872/j.issn.1007-385X.2013.03.006
Abstract:
Objective:To investigate the expression of miRNA-210(miR-210) in breast cancer tissues and its effect on proliferation, migration and invasion of breast cancer MDA-MB-231 cells. Methods: Tissues of breast cancer patients were collected from Department of Medical Oncology, First Affiliated Hospital of Kunming Medical University during October 2011 to June 2012. The expressions of miR-210 were compared between breast cancer tissues and the para-carcinoma tissues of 20 patients, as well as between breast cancer MDA-MB-231 cells and normal breast MCF-10a cells by real-time PCR. miR-210 inhibitor was transfected into breast cancer MDA-MB-231 cells by LipofectamineTM 2000 and the transfection efficiency was examined under a fluorescence microscope. Cell proliferation was evaluated by MTT assay and soft-agar colony formation assay. The cell cycle and apoptosis were detected by flow cytometry assay. The cell migration and invasion abilities were detected by migration and invasion assay. Results: The expressions of miR-210 in breast cancer tissues and cells were both significantly higher than those in para-carcinoma tissues and normal breast cells (P<0.01). miR-210 inhibitor was successfully transfected into MDA-MB-231 cells with a high transfection efficiency of (88.29±2.98)%. The proliferation ability of MDA-MB-231 cells was decreased significantly after transfection of miR-210 inhibitor (P<0.05). The percentages of cells in G0/G1 phase (\[64.23±3.12\]% vs \[5553±0.96\]%, P<0.01\] and of the apoptotic cells (\[31.90±305\]% vs \[15.98±0.63\]%, P<0.01） were significantly increased. The migration (\[291.00±43.12\] vs \[137.38±8349\], P<0.01\] and invasion (\[131.63±32.01\] vs \[647.88±31.20\], P<0.01\] of MDA-MB-231 cells were significantly inhibited. Conclusion:miR-210 is over-expressed in breast cancer tissues and cells. The proliferation, migration and invasion of human breast cancer MDA-MB-231 cells are inhibited after the transfection of miR-210 inhibitor.
Objective:To investigate the expression of miRNA-210(miR-210) in breast cancer tissues and its effect on proliferation, migration and invasion of breast cancer MDA-MB-231 cells. Methods: Tissues of breast cancer patients were collected from Department of Medical Oncology, First Affiliated Hospital of Kunming Medical University during October 2011 to June 2012. The expressions of miR-210 were compared between breast cancer tissues and the para-carcinoma tissues of 20 patients, as well as between breast cancer MDA-MB-231 cells and normal breast MCF-10a cells by real-time PCR. miR-210 inhibitor was transfected into breast cancer MDA-MB-231 cells by LipofectamineTM 2000 and the transfection efficiency was examined under a fluorescence microscope. Cell proliferation was evaluated by MTT assay and soft-agar colony formation assay. The cell cycle and apoptosis were detected by flow cytometry assay. The cell migration and invasion abilities were detected by migration and invasion assay. Results: The expressions of miR-210 in breast cancer tissues and cells were both significantly higher than those in para-carcinoma tissues and normal breast cells (P<0.01). miR-210 inhibitor was successfully transfected into MDA-MB-231 cells with a high transfection efficiency of (88.29±2.98)%. The proliferation ability of MDA-MB-231 cells was decreased significantly after transfection of miR-210 inhibitor (P<0.05). The percentages of cells in G0/G1 phase (\[64.23±3.12\]% vs \[5553±0.96\]%, P<0.01\] and of the apoptotic cells (\[31.90±305\]% vs \[15.98±0.63\]%, P<0.01） were significantly increased. The migration (\[291.00±43.12\] vs \[137.38±8349\], P<0.01\] and invasion (\[131.63±32.01\] vs \[647.88±31.20\], P<0.01\] of MDA-MB-231 cells were significantly inhibited. Conclusion:miR-210 is over-expressed in breast cancer tissues and cells. The proliferation, migration and invasion of human breast cancer MDA-MB-231 cells are inhibited after the transfection of miR-210 inhibitor.
2013, 20(3):295-300.
DOI: 10.3872/j.issn.1007-385X.2013.03.007
Abstract:
Objective:To construct a recombinant eukaryotic expression vector encoding miRNA-126 and to explore its effect on proliferation and migration of mouse breast cancer 4T1 cells. Methods: Sense and antisense oligonucleotides of miRNA-126 were designed and synthesized respectively. The eukaryotic expression vector pcDNA6.2-miR-126 was constructed and transiently transfected into 4T1 cells in vitro. The transfection efficiency was observed under a fluorescent microscope. The expression level of miRNA-126 in 4T1 cells was determined by real-time PCR. The proliferation and colony formation ability of 4T1 cells were detected by MTT assay and colony formation assay. The migration of 4T1 cells in vitro was determined by scratch assay. Results: pcDNA62-miR-126 eukaryotic expression vector was successfully constructed and miRNA-126 was effectively expressed in 4T1 cells. Compared with that in empty plasmid transfected group (pcDNA6.2-Ctrl) , the proliferation capacity of 4T1 cells in vitro was obviously decreased in pcDNA6.2-miR-126 transfected group after transient transfection for 72 hours （\[0.30±0.03\] vs \[0.51±0.04\], P<0.05). The migration capacity of 4T1 cells in pcDNA6.2-miR-126 transfected group was also significantly inhibited after transfection for 48 hours (\[8.17±2.30\] vs \[28.33±2.16\], P<0.05). Conclusion:Overexpression of miRNA-126 may inhibit the proliferation and migration of breast cancer 4T1 cells.
Objective:To construct a recombinant eukaryotic expression vector encoding miRNA-126 and to explore its effect on proliferation and migration of mouse breast cancer 4T1 cells. Methods: Sense and antisense oligonucleotides of miRNA-126 were designed and synthesized respectively. The eukaryotic expression vector pcDNA6.2-miR-126 was constructed and transiently transfected into 4T1 cells in vitro. The transfection efficiency was observed under a fluorescent microscope. The expression level of miRNA-126 in 4T1 cells was determined by real-time PCR. The proliferation and colony formation ability of 4T1 cells were detected by MTT assay and colony formation assay. The migration of 4T1 cells in vitro was determined by scratch assay. Results: pcDNA62-miR-126 eukaryotic expression vector was successfully constructed and miRNA-126 was effectively expressed in 4T1 cells. Compared with that in empty plasmid transfected group (pcDNA6.2-Ctrl) , the proliferation capacity of 4T1 cells in vitro was obviously decreased in pcDNA6.2-miR-126 transfected group after transient transfection for 72 hours （\[0.30±0.03\] vs \[0.51±0.04\], P<0.05). The migration capacity of 4T1 cells in pcDNA6.2-miR-126 transfected group was also significantly inhibited after transfection for 48 hours (\[8.17±2.30\] vs \[28.33±2.16\], P<0.05). Conclusion:Overexpression of miRNA-126 may inhibit the proliferation and migration of breast cancer 4T1 cells.
2013, 20(3):301-305.
DOI: 10.3872/j.issn.1007-385X.2013.03.008
Abstract:
Objective:To investigate the effect of in vitro silencing Kruppel like factor 4 ( KLF4 ) gene expression on the proliferation and migration of esophageal cancer KYSE140 cells. Methods: Western blotting was used to detect the expression of KLF4 protein in the esophageal cancer cell lines, including KYSE140, KYSE150, EC109 and EC9706，and immortalized esophageal NE3 cells. Two pairs of siRNAs targeting KLF4 (KLF4-siRNA1，KLF4-siRNA2) and control siRNA (Ctrl-siRNA) were chemically synthesized, and were transfected into KYSE140 cells with a high expression of KLF4 in vitro to form KLF4-siRNA1-KYSE140, KLF4-siRNA2-KYSE140 and Ctrl-siRNA-KYSE140 cells. The proliferation and migration of esophageal cancer KYSE140 cells after transfection were detected by MTT assay and Transwell assay, respectively. Results: The expression of KLF4 in KYSE140 cells was higher than that in KYSE150, EC109 and EC9706 cells (\[562±0.02\] vs \[171±0.23\], \[3.24±0.35\], \[3.16±0.41\], both P<0.05). Compared with that in Ctrl-siRNA-KYSE140 cells, the expression of KLF4 protein was significantly decreased in KLF4-siRNA1-KYSE140 and KLF4-siRNA2-KYSE140 cells (\[0.49±0.18\], \[0.32±0.09\] vs \[0.98±0.19\], both P＜0.05), the capacities of proliferation were significantly increased (\[12±0.8\], \[1.4±0.1\] vs \[0.6±0.1\], both P<0.05), and the numbers of migration were also significantly increased (\[780±22\], \[475±25\] vs \[83±17\], P<0.05). Conclusion:KLF4 functions as a negative regulator in the proliferation and migration of esophageal cancer cells.
Objective:To investigate the effect of in vitro silencing Kruppel like factor 4 ( KLF4 ) gene expression on the proliferation and migration of esophageal cancer KYSE140 cells. Methods: Western blotting was used to detect the expression of KLF4 protein in the esophageal cancer cell lines, including KYSE140, KYSE150, EC109 and EC9706，and immortalized esophageal NE3 cells. Two pairs of siRNAs targeting KLF4 (KLF4-siRNA1，KLF4-siRNA2) and control siRNA (Ctrl-siRNA) were chemically synthesized, and were transfected into KYSE140 cells with a high expression of KLF4 in vitro to form KLF4-siRNA1-KYSE140, KLF4-siRNA2-KYSE140 and Ctrl-siRNA-KYSE140 cells. The proliferation and migration of esophageal cancer KYSE140 cells after transfection were detected by MTT assay and Transwell assay, respectively. Results: The expression of KLF4 in KYSE140 cells was higher than that in KYSE150, EC109 and EC9706 cells (\[562±0.02\] vs \[171±0.23\], \[3.24±0.35\], \[3.16±0.41\], both P<0.05). Compared with that in Ctrl-siRNA-KYSE140 cells, the expression of KLF4 protein was significantly decreased in KLF4-siRNA1-KYSE140 and KLF4-siRNA2-KYSE140 cells (\[0.49±0.18\], \[0.32±0.09\] vs \[0.98±0.19\], both P＜0.05), the capacities of proliferation were significantly increased (\[12±0.8\], \[1.4±0.1\] vs \[0.6±0.1\], both P<0.05), and the numbers of migration were also significantly increased (\[780±22\], \[475±25\] vs \[83±17\], P<0.05). Conclusion:KLF4 functions as a negative regulator in the proliferation and migration of esophageal cancer cells.
9 Silencing AQP-5 on proliferation, apoptosis and chemosensitivity of human colon cancer HT-29 cells
2013, 20(3):306-311.
DOI: 10.3872/j.issn.1007-385X.2013.03.009
Abstract:
Objective:To investigate the effect of siRNA (small interference RNA, siRNA) silencing aquaporin-5 ( AQP-5 ) expression on the proliferation, apoptosis, and chemosensitivity of human colon cancer HT-29 cells. Methods: A synthetic AQP-5-siRNA sequence was transfected into HT-29 cells, and the inhibition efficiency was detected by Western blotting. Sulphorhodamine B (SRB) assay were used to detect the cell proliferation inhibition rate. Cell apoptosis of HT-29 cells were detected by flow cytometry (FCM). The activity of caspase-3 in HT-29 cells was measured by spectrophotometry. The mRNA and protein levels of PCNA and P53 in HT-29 cells after AQP-5-siRNA transfection were determined by real-time PCR and Western blotting. 5-fluorouracil (5-FU) and cisplatin (DDP) were selected to treat the AQP-5-siRNA-HT-29 cells, and SRB assay was used to detect the cell proliferation inhibition rate. The “Q” Method of Jin Zhenjun was used to evaluate the interaction of the two drugs. Results: SRB assay showed that compared with NC-HT-29 cells, AQP-5 expression was significantly decreased in AQP-5-siRNA-HT-29 cells (P<0.05). Compared with NC-HT-29 cells，the proliferation inhibition rate was increased significantly in AQP-5-siRNA-HT-29 cells (\[9.23±051\]% vs 0, P<0.05). Flow cytometry analysis showed that compared with NC-HT-29 cells, the cell apoptosis rate increased dramatically in AQP-5-siRNA-HT-29 cells (\[10.81±1.32\]% vs \[0.99±0.18\]%, P<0.05). The activities of caspase-3 were also increased in AQP-5-siRNA-HT-29 cells compared with NC-HT-29 cells (\[0.19±0.03\] vs \[0.09±0.01\], P<0.05). Real-time PCR and Western blotting results indicated that compared with NC-HT-29 cells, the mRNA and protein levels of PCNA were decreased (P<0.05), simultaneously, the mRNA and protein levels of P53 were increased (P<0.05) in AQP-5-siRNA-HT-29 cells. Compared with AQP-5-siRNA-HT-29 cells or 5-FU-HT-29 cells, the proliferation inhibition rate was increased remarkably in AQP-5-siRNA+5-FU-HT-29 cells(\[44.93±2.28\]% vs \[9.11±032\]%, \[25.68±171\]%, P<0.05), respectively. Compared with AQP-5-siRNA-HT-29 cells or DDP-HT-29 cells, the proliferation inhibition rate was increased remarkably in AQP-5-siRNA+DDP-HT-29 cells (\[39.01±176\]% vs \[911±0.32\]%, \[18.47±1.25\]%, P<0.05), respectively. Moreover, the synergistic effects were showed between AQP-5-siRNA and 5-FU or DDP with Q value of 1.38 and 1.51, respectively. Conclusion:AQP-5-siRNA can inhibit the proliferation, promote the apoptosis of HT-29 cells, and increase the chemosensitivity of HT-29 cells to 5-FU and DDP.
Objective:To investigate the effect of siRNA (small interference RNA, siRNA) silencing aquaporin-5 ( AQP-5 ) expression on the proliferation, apoptosis, and chemosensitivity of human colon cancer HT-29 cells. Methods: A synthetic AQP-5-siRNA sequence was transfected into HT-29 cells, and the inhibition efficiency was detected by Western blotting. Sulphorhodamine B (SRB) assay were used to detect the cell proliferation inhibition rate. Cell apoptosis of HT-29 cells were detected by flow cytometry (FCM). The activity of caspase-3 in HT-29 cells was measured by spectrophotometry. The mRNA and protein levels of PCNA and P53 in HT-29 cells after AQP-5-siRNA transfection were determined by real-time PCR and Western blotting. 5-fluorouracil (5-FU) and cisplatin (DDP) were selected to treat the AQP-5-siRNA-HT-29 cells, and SRB assay was used to detect the cell proliferation inhibition rate. The “Q” Method of Jin Zhenjun was used to evaluate the interaction of the two drugs. Results: SRB assay showed that compared with NC-HT-29 cells, AQP-5 expression was significantly decreased in AQP-5-siRNA-HT-29 cells (P<0.05). Compared with NC-HT-29 cells，the proliferation inhibition rate was increased significantly in AQP-5-siRNA-HT-29 cells (\[9.23±051\]% vs 0, P<0.05). Flow cytometry analysis showed that compared with NC-HT-29 cells, the cell apoptosis rate increased dramatically in AQP-5-siRNA-HT-29 cells (\[10.81±1.32\]% vs \[0.99±0.18\]%, P<0.05). The activities of caspase-3 were also increased in AQP-5-siRNA-HT-29 cells compared with NC-HT-29 cells (\[0.19±0.03\] vs \[0.09±0.01\], P<0.05). Real-time PCR and Western blotting results indicated that compared with NC-HT-29 cells, the mRNA and protein levels of PCNA were decreased (P<0.05), simultaneously, the mRNA and protein levels of P53 were increased (P<0.05) in AQP-5-siRNA-HT-29 cells. Compared with AQP-5-siRNA-HT-29 cells or 5-FU-HT-29 cells, the proliferation inhibition rate was increased remarkably in AQP-5-siRNA+5-FU-HT-29 cells(\[44.93±2.28\]% vs \[9.11±032\]%, \[25.68±171\]%, P<0.05), respectively. Compared with AQP-5-siRNA-HT-29 cells or DDP-HT-29 cells, the proliferation inhibition rate was increased remarkably in AQP-5-siRNA+DDP-HT-29 cells (\[39.01±176\]% vs \[911±0.32\]%, \[18.47±1.25\]%, P<0.05), respectively. Moreover, the synergistic effects were showed between AQP-5-siRNA and 5-FU or DDP with Q value of 1.38 and 1.51, respectively. Conclusion:AQP-5-siRNA can inhibit the proliferation, promote the apoptosis of HT-29 cells, and increase the chemosensitivity of HT-29 cells to 5-FU and DDP.
Liu Zhipeng
,
Wang Zonghua
,
Lu Bin
,
Hu Chunyan
,
Li Xuecheng
,
Zou Liquan
,
Zhang Fangzheng
,
Chen Ling
2013, 20(3):312-316.
DOI: 10.3872/j.issn.1007-385X.2013.03.010
Abstract:
Objective:To explore the inhibitory effect of miRNA-29a(miR-29a) on the proliferation of human gastric cancer cell lines SGC-7901 and AGS through over-expression of miR-29a mediated by the recombinant replication-deficient human adenovirus type 5 vector. Methods: The recombinant adenovirus Ad-miR-29a containing pre-miR-29a or control adenovirus Ad-LacZ containing LacZ gene was constructed and infected into human gastric cancer SGC-7901 and AGS cells, respectively. The expressions of miR-29a in SGC-7901 and AGS cells were detected by real-time PCR. CCK-8 assay was employed to examine the inhibitory effect of miR-29a on the proliferation of human gastric cancer cell lines. The flow cytometry assay was used to analyze cell cycle of SGC-7901 and AGS cells. The migration ablilities of SGC-7901 and AGS cells were assessed by Transwell assay. Results: Compared with the Ad-LacZ group, the Ad-miR29a group expressed higher level of miR-29a in both SGC-7901 and AGS cells (\[17.35±0.71\] vs \[1.12±0.09\], \[26.50±109\] vs \[0.95±0.04\], P<001). The proliferation of SGC-7901 and AGS was significantly inhibited in the Ad-miR29a group as compared with that in the Ad-LacZ group. Six days after Ad-miR29a infection, the D value was significantly decreased (\[0.54± 003\] vs \[0.77 ± 0.04\], \[0.70 ± 0.03\] vs \[0.88 ± 0.04\], P<0.01). The proliferation of cells in the Ad-LacZ group showed no significant changes as compared with that in the uninfected group (P>005). Meanwhile, The percentage of SGC-7901 or AGS cells arrested in G0/G1 period in the Ad-miR29a group was significantly higher than that in the Ad-LacZ group (\[63.10±4.91\]% vs \[47.60±5.31\]%，\[69.80±3.15\]% vs \[5460±422\]%, P<0.05\]. However, no significant changes were found in the migration ability of gastric cancer SGC-7901 and AGS cells. Conclusion:miR-29a can effectively suppress the proliferation of human gastric cancer cells, which makes a promising new therapeutic target for gastric cancer.
Objective:To explore the inhibitory effect of miRNA-29a(miR-29a) on the proliferation of human gastric cancer cell lines SGC-7901 and AGS through over-expression of miR-29a mediated by the recombinant replication-deficient human adenovirus type 5 vector. Methods: The recombinant adenovirus Ad-miR-29a containing pre-miR-29a or control adenovirus Ad-LacZ containing LacZ gene was constructed and infected into human gastric cancer SGC-7901 and AGS cells, respectively. The expressions of miR-29a in SGC-7901 and AGS cells were detected by real-time PCR. CCK-8 assay was employed to examine the inhibitory effect of miR-29a on the proliferation of human gastric cancer cell lines. The flow cytometry assay was used to analyze cell cycle of SGC-7901 and AGS cells. The migration ablilities of SGC-7901 and AGS cells were assessed by Transwell assay. Results: Compared with the Ad-LacZ group, the Ad-miR29a group expressed higher level of miR-29a in both SGC-7901 and AGS cells (\[17.35±0.71\] vs \[1.12±0.09\], \[26.50±109\] vs \[0.95±0.04\], P<001). The proliferation of SGC-7901 and AGS was significantly inhibited in the Ad-miR29a group as compared with that in the Ad-LacZ group. Six days after Ad-miR29a infection, the D value was significantly decreased (\[0.54± 003\] vs \[0.77 ± 0.04\], \[0.70 ± 0.03\] vs \[0.88 ± 0.04\], P<0.01). The proliferation of cells in the Ad-LacZ group showed no significant changes as compared with that in the uninfected group (P>005). Meanwhile, The percentage of SGC-7901 or AGS cells arrested in G0/G1 period in the Ad-miR29a group was significantly higher than that in the Ad-LacZ group (\[63.10±4.91\]% vs \[47.60±5.31\]%，\[69.80±3.15\]% vs \[5460±422\]%, P<0.05\]. However, no significant changes were found in the migration ability of gastric cancer SGC-7901 and AGS cells. Conclusion:miR-29a can effectively suppress the proliferation of human gastric cancer cells, which makes a promising new therapeutic target for gastric cancer.
2013, 20(3):317-322.
DOI: 10.3872/j.issn.1007-385X.2013.03.011
Abstract:
Objective:To investigate the expression and aberrant methylation of growth arrest and DNA-damage-inducible 45 alpha ( GADD45A ) gene in gastric cardia adenocarcinoma (GCA) and to explore its clinical significance. Methods: Tissue samples in GCA patients (138 cases) were selected from Fourth Hospital of Hebei University during 2004 to 2007. Bisulfite sequencing (BS-Seq), bisulfite conversion-methylation specific polymerase chain reaction (BS-MSP), reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry methods were used respectively to detect the methylation status, mRNA and protein expression of GADD45A gene in GCA tissues and the para-carcinoma normal tissues. Results: The methylation frequency of four CpG sites in distal promoter of GADD45A in GCA tissues (44.93%, \[62/138\]) was significantly higher than that in the para-carcinoma normal tissues (0.00%, 0/138) (P<0.01). The methylation frequency of 4 CpG sites in stage Ⅲ and Ⅳ GCA tissues was significantly higher than that in stage Ⅰ and Ⅱ GCA tissues (P<0.05). However, the methylation status GADD45A in GCA tissues was not correltaed with age, gender and pathological differentiation (P>0.05). For GADD45A region 2 and 3 located in proximal promoter and exon 1, no methylation was detected in GCA and the para-carcinoma normal tissues. The expression of GADD45A mRNA and positive expression rate of GADD45A protein in GCA tissues were significantly lower than those in the para-carcinoma normal tissues (\[0.35±0.15\] vs \[0.78±0.26\], 42.75% vs 71.01%, P<0.05) and was associated with methylation status of 4 CpG sites in distal promoter (r=-0.52, P<0.01). Conclusion:Hypermethylation of four CpG sites in distal promoter of GADD45A gene may be responsible for the decreased expression of GADD45A in GCA.
Objective:To investigate the expression and aberrant methylation of growth arrest and DNA-damage-inducible 45 alpha ( GADD45A ) gene in gastric cardia adenocarcinoma (GCA) and to explore its clinical significance. Methods: Tissue samples in GCA patients (138 cases) were selected from Fourth Hospital of Hebei University during 2004 to 2007. Bisulfite sequencing (BS-Seq), bisulfite conversion-methylation specific polymerase chain reaction (BS-MSP), reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry methods were used respectively to detect the methylation status, mRNA and protein expression of GADD45A gene in GCA tissues and the para-carcinoma normal tissues. Results: The methylation frequency of four CpG sites in distal promoter of GADD45A in GCA tissues (44.93%, \[62/138\]) was significantly higher than that in the para-carcinoma normal tissues (0.00%, 0/138) (P<0.01). The methylation frequency of 4 CpG sites in stage Ⅲ and Ⅳ GCA tissues was significantly higher than that in stage Ⅰ and Ⅱ GCA tissues (P<0.05). However, the methylation status GADD45A in GCA tissues was not correltaed with age, gender and pathological differentiation (P>0.05). For GADD45A region 2 and 3 located in proximal promoter and exon 1, no methylation was detected in GCA and the para-carcinoma normal tissues. The expression of GADD45A mRNA and positive expression rate of GADD45A protein in GCA tissues were significantly lower than those in the para-carcinoma normal tissues (\[0.35±0.15\] vs \[0.78±0.26\], 42.75% vs 71.01%, P<0.05) and was associated with methylation status of 4 CpG sites in distal promoter (r=-0.52, P<0.01). Conclusion:Hypermethylation of four CpG sites in distal promoter of GADD45A gene may be responsible for the decreased expression of GADD45A in GCA.
12 Expression of ubiquitin-conjugating enzyme H10 in meningioma tissues and its clinical significance
2013, 20(3):323-329.
DOI: 10.3872/j.issn.1007-385X.2013.03.012
Abstract:
Objective:To investigate the expression of ubiquitin-conjugating enzyme H10 (UbcH10) protein in meningiomas tissues of different pathological grades and its clinical significance. 〖WTHZ〗Methods：〖WTBZ〗Forty-seven patients diagnosed as meningioma with integrity clinical and follow-up information during April 2002 to September 2009 were selected from Department of Neurosurgery, Changzheng Hospital and were graded according to the 2007 WHO standard. The expressions of UbcH10 and Ki-67 proteins in meningioma tissues of various pathological grades were examined by immunohistochemistry. The correlation between the expressions of UbcH10 and Ki-67 protein was analyzed by using Spearman method. The correlation of UbcH10 expression with meningioma recurrence-free surivival was analyzed using the Kaplan-Meier method. 〖WTHZ〗Results：〖WTBZ〗UbcH10 protein was mainly expressed in the cytoplasm of meningioma cells. The expression of UbcH10 protein in meningioma tissues was significantly higher than that in normal brain tissues (\[5.57±1.72\]% vs 0，P=0.00). Quantitative analysis confirmed that the expression of UbcH10 protein was significantly higher in atypical and anaplastic meningiomas as compared with classic meningiomas (\[10.53±5.79\]% vs \[4.23±2.85\]%, P<0.01), indicating that the expression of UbcH10 protein was correlated with meningioma pathological grade and recurrence. The expression of UbcH10 protein was correlated with the expression of Ki-67 protein (r=0.77, P<0.01). Kaplan-Meier survival curve result indicated patients with high expression level of UbcH10 and Ki-67 and higher pathological grade showed a significantly shortened recurrence-free surivival than those with a lower expression level of UbcH10 (P=0.007) and Ki-67 (P=0.018), and a lower pathological grade (P<0.01). Cox multivariate regression analysis showed that the pathological grade (P=0.029) and the expression level of UbcH10 (P=0.044) were independent prognosis factors for meningioma patients, with the hazard ratio of 2.918 and 4756, respectively. Whereas, the clinicopathological factors, including age, gender, Ki-67 level, tumor diameter and tumor location were not independent factors for prognosis. Conclusion:UbcH10 may be involved in the development and progression of meningioma, and UbcH10 expression is an independent risk factor that could predict the prognosis.
Objective:To investigate the expression of ubiquitin-conjugating enzyme H10 (UbcH10) protein in meningiomas tissues of different pathological grades and its clinical significance. 〖WTHZ〗Methods：〖WTBZ〗Forty-seven patients diagnosed as meningioma with integrity clinical and follow-up information during April 2002 to September 2009 were selected from Department of Neurosurgery, Changzheng Hospital and were graded according to the 2007 WHO standard. The expressions of UbcH10 and Ki-67 proteins in meningioma tissues of various pathological grades were examined by immunohistochemistry. The correlation between the expressions of UbcH10 and Ki-67 protein was analyzed by using Spearman method. The correlation of UbcH10 expression with meningioma recurrence-free surivival was analyzed using the Kaplan-Meier method. 〖WTHZ〗Results：〖WTBZ〗UbcH10 protein was mainly expressed in the cytoplasm of meningioma cells. The expression of UbcH10 protein in meningioma tissues was significantly higher than that in normal brain tissues (\[5.57±1.72\]% vs 0，P=0.00). Quantitative analysis confirmed that the expression of UbcH10 protein was significantly higher in atypical and anaplastic meningiomas as compared with classic meningiomas (\[10.53±5.79\]% vs \[4.23±2.85\]%, P<0.01), indicating that the expression of UbcH10 protein was correlated with meningioma pathological grade and recurrence. The expression of UbcH10 protein was correlated with the expression of Ki-67 protein (r=0.77, P<0.01). Kaplan-Meier survival curve result indicated patients with high expression level of UbcH10 and Ki-67 and higher pathological grade showed a significantly shortened recurrence-free surivival than those with a lower expression level of UbcH10 (P=0.007) and Ki-67 (P=0.018), and a lower pathological grade (P<0.01). Cox multivariate regression analysis showed that the pathological grade (P=0.029) and the expression level of UbcH10 (P=0.044) were independent prognosis factors for meningioma patients, with the hazard ratio of 2.918 and 4756, respectively. Whereas, the clinicopathological factors, including age, gender, Ki-67 level, tumor diameter and tumor location were not independent factors for prognosis. Conclusion:UbcH10 may be involved in the development and progression of meningioma, and UbcH10 expression is an independent risk factor that could predict the prognosis.
2013, 20(3):330-335.
DOI: 10.3872/j.issn.1007-385X.2013.03.013
Abstract:
Objective:To explore the expressions of receptors before and after NK cell amplification from patients with esophageal cancer and its cytotoxicity to tumor cells. Methods: Peripheral blood was collected from the Fujian Provincial Tumor Hospital, including 20 cases of esophageal cancer patients and 10 cases of healthy donors (control group). NK cells were amplified by combination of IL-2+IL-12+IL-15+IL-18. Cell immunophenotype and NK cell receptor expressions (CD56+, CD69+，NKG2D, NKp30, NKp44, NKp46, CD158b and CD159a) were determined by flow cytometry. The cytotoxicity of NK cells to various tumor cell lines (K562, Raji, Eca-109 and TE-1) were detected by lactate dehydrogenase (LDH) assay. Results: Compared with the control group, the ratio of CD3+, CD4+and CD4+/CD8+T cells were significantly lower in the peripheral blood of patients with esophageal cancer (P<0.05), and the ratios of NK cells (CD56+) and regulatory T cells (Treg) were significantly higher (P<0.05).The ratio of NK cells (CD3-CD56+) increased up to 90% after being cultured in combination of IL-2+IL-12+IL-15+IL-18 for 20 days. NK cell count expanded up to 1 000 times (P<0.01). The ratios of CD3+, CD4+T cells, B cells (CD19), monocyte-macrophage cells (CD14) and regulatory T cells (T-reg) were in a significant reduction 20 days after culture (P<0.01), with no significant difference between the patients with esophageal cancer and the control group (P>0.05). CD69+and NK cell activating receptors (NKG2D, NKp30, NKp44, NKp46) were significantly increased and the inhibitory receptors (CD158b, CD159a) were significantly decreased (P<0.05). NK cells derived from patients with esophageal cancer showed significant increase of cytotoxicity on tumor cells K562, Raji, Eca-109 and TE-1 after culture for 20 days compared to those before culture (\[69.2±51\]% vs \[42.3±30\]%, \[44.6±3.2\]% vs \[21.1±2.0\]%, \[69.7±3.9\]% vs \[50.3±3.5\]%, \[67.1±4.5\]% vs \[41.2±3.3\]%, P<0.01). Conclusion:The combined cytokines of IL-2+IL-12+IL-15+IL-18 can effectively expand peripheral blood NK cells, up-regulate the expressions of activated receptor and down-regulate the expression of inhibitory receptors. The numbers and functions of the cultured NK cells can both meet the clinical treatment needs.
Objective:To explore the expressions of receptors before and after NK cell amplification from patients with esophageal cancer and its cytotoxicity to tumor cells. Methods: Peripheral blood was collected from the Fujian Provincial Tumor Hospital, including 20 cases of esophageal cancer patients and 10 cases of healthy donors (control group). NK cells were amplified by combination of IL-2+IL-12+IL-15+IL-18. Cell immunophenotype and NK cell receptor expressions (CD56+, CD69+，NKG2D, NKp30, NKp44, NKp46, CD158b and CD159a) were determined by flow cytometry. The cytotoxicity of NK cells to various tumor cell lines (K562, Raji, Eca-109 and TE-1) were detected by lactate dehydrogenase (LDH) assay. Results: Compared with the control group, the ratio of CD3+, CD4+and CD4+/CD8+T cells were significantly lower in the peripheral blood of patients with esophageal cancer (P<0.05), and the ratios of NK cells (CD56+) and regulatory T cells (Treg) were significantly higher (P<0.05).The ratio of NK cells (CD3-CD56+) increased up to 90% after being cultured in combination of IL-2+IL-12+IL-15+IL-18 for 20 days. NK cell count expanded up to 1 000 times (P<0.01). The ratios of CD3+, CD4+T cells, B cells (CD19), monocyte-macrophage cells (CD14) and regulatory T cells (T-reg) were in a significant reduction 20 days after culture (P<0.01), with no significant difference between the patients with esophageal cancer and the control group (P>0.05). CD69+and NK cell activating receptors (NKG2D, NKp30, NKp44, NKp46) were significantly increased and the inhibitory receptors (CD158b, CD159a) were significantly decreased (P<0.05). NK cells derived from patients with esophageal cancer showed significant increase of cytotoxicity on tumor cells K562, Raji, Eca-109 and TE-1 after culture for 20 days compared to those before culture (\[69.2±51\]% vs \[42.3±30\]%, \[44.6±3.2\]% vs \[21.1±2.0\]%, \[69.7±3.9\]% vs \[50.3±3.5\]%, \[67.1±4.5\]% vs \[41.2±3.3\]%, P<0.01). Conclusion:The combined cytokines of IL-2+IL-12+IL-15+IL-18 can effectively expand peripheral blood NK cells, up-regulate the expressions of activated receptor and down-regulate the expression of inhibitory receptors. The numbers and functions of the cultured NK cells can both meet the clinical treatment needs.
Wang Xiaomeng
,
Li Ling
,
Yu Jinpu
,
Li Hui
,
Qi Jing
,
Zhang Peng
,
Yu Wenwen
,
Ren Xiubao
,
Cao Shui
2013, 20(3):336-341.
DOI: 10.3872/j.issn.1007-385X.2013.03.014
Abstract:
Objective:By comparing the cell immunophenotype, the expansion fold and the cytotoxic activity of the expansion products in various cell culture methods to identify a more effective solution for the in vitro expansion of NK cells. Methods: Four methods for expansion of NK cells from peripheral blood were established, including method one, a classical culture protocol for NK cells (IL-2+IL-15), method two (IL-2+IL-15+IL- 18), method three (IL-2+IL-15+IL-7), and method four, a novel NK-specific culture medium (IL- 2+OKT3). 10 patients with advanced solid tumors in Department of Biotherapy, Cancer Hospital Affiliated to Tianjin Medical University from February 2012 to April 2012 were obtained, and PBMCs from those patients were isolated by Ficoll-Hypaque density gradient centrifugation. The proportion of different lymphocyte subsets (especially for NK cells) were detected by flow cytometry on 0, 5, 10 and 15 days. The changes of NK cell expansion fold and the proportion of different lymphocyte subsets were detected among 4 groups after expansion in vitro for 15 days. The anti-tumor ytotoxicity against human K562 cell line among 4 groups were measured using LDH assay. Results: After expansion for 15 days, the expansion folds of total cells in 4 groups were (40.1±2000), (44.08±22.09), (44.82±23.67) and (46.82±25.02), respectively. The proportion of NK cells in 4 groups increased from (20.44±2.23)% on day 0 to (48.30±13.90)%, (54.72±12.25)%, (55.94±12.70)% and (54.5±14.93)% on day 15, respectively. The expansion folds of NK cells in 4 groups were (75.86±28.57), (93.32±32.16), (88.66±24.94) and (58.88±4153), respectively. No significant difference was found on the total cell expansion, NK cell expansion folds among the 4 groups (P<0.05). The cytotoxic activity of the expansion products in methods one, two and three were higher than that of method four in vitro (\[63.40±5.00\]%, \[77.30±940\]%, [62.17±5.60\]% vs \[37.39±10.42\]%，P<0.05). There was no significant difference among the first 3 groups (P >0.05). Conclusion:NK-specific cytokines have great influence on the expansion of NK cells in vitro. However, no significant difference is found among various cytokine combinations. The cytotoxic activity of the expansion products in methods one, two and three against K562 cells are significantly higher than that of method four.
Objective:By comparing the cell immunophenotype, the expansion fold and the cytotoxic activity of the expansion products in various cell culture methods to identify a more effective solution for the in vitro expansion of NK cells. Methods: Four methods for expansion of NK cells from peripheral blood were established, including method one, a classical culture protocol for NK cells (IL-2+IL-15), method two (IL-2+IL-15+IL- 18), method three (IL-2+IL-15+IL-7), and method four, a novel NK-specific culture medium (IL- 2+OKT3). 10 patients with advanced solid tumors in Department of Biotherapy, Cancer Hospital Affiliated to Tianjin Medical University from February 2012 to April 2012 were obtained, and PBMCs from those patients were isolated by Ficoll-Hypaque density gradient centrifugation. The proportion of different lymphocyte subsets (especially for NK cells) were detected by flow cytometry on 0, 5, 10 and 15 days. The changes of NK cell expansion fold and the proportion of different lymphocyte subsets were detected among 4 groups after expansion in vitro for 15 days. The anti-tumor ytotoxicity against human K562 cell line among 4 groups were measured using LDH assay. Results: After expansion for 15 days, the expansion folds of total cells in 4 groups were (40.1±2000), (44.08±22.09), (44.82±23.67) and (46.82±25.02), respectively. The proportion of NK cells in 4 groups increased from (20.44±2.23)% on day 0 to (48.30±13.90)%, (54.72±12.25)%, (55.94±12.70)% and (54.5±14.93)% on day 15, respectively. The expansion folds of NK cells in 4 groups were (75.86±28.57), (93.32±32.16), (88.66±24.94) and (58.88±4153), respectively. No significant difference was found on the total cell expansion, NK cell expansion folds among the 4 groups (P<0.05). The cytotoxic activity of the expansion products in methods one, two and three were higher than that of method four in vitro (\[63.40±5.00\]%, \[77.30±940\]%, [62.17±5.60\]% vs \[37.39±10.42\]%，P<0.05). There was no significant difference among the first 3 groups (P >0.05). Conclusion:NK-specific cytokines have great influence on the expansion of NK cells in vitro. However, no significant difference is found among various cytokine combinations. The cytotoxic activity of the expansion products in methods one, two and three against K562 cells are significantly higher than that of method four.
Wang Xiaoyan
,
Wang Xiang
,
Zhao Linghui
,
Zheng Yukun
,
Chen Jingxian
,
Shen Chongyang
,
Deng Hongmei
,
Zhang Xiaotong
,
Qi Julan
,
Zhang Rong
,
Hai Quan
2013, 20(3):342-347.
DOI: 10.3872/j.issn.1007-385X.2013.03.015
Abstract:
Objective:To observe the effect of in vitro culture of different time on proliferation, immunophenotype, cytotoxicity and cytokine secretion of human cord blood derived cytokine induced killer (CIK) cells after cryopreservation.Methods: CIK cells were cryopreserved for 1 month after in vitro culture for 6, 9 and 12 d. The cell proliferation was detected every 24 h when they were recovered and cultured for 72 h. The immunophenotype, cytotoxicity on A549 cells and the secretion of IFN-γ and TNF-α of CIK cells after recovery were detected by flow cytometry, CCK-8 assay and ELISA assay, respectively. Results:CIK cells, in vitro culture of different time before cryopreservation still showed a superior proliferation activity after recovery, among which the proliferation of CIK cells in vitro cultured for 6 d before cryopreservation, was significantly higher than those in vitro cultured for 6 and 12 d before cryopreservation. The CIK cell counts were (\[35.90±1.67\]×106, \[18.98±2.13\]×106, and \[11.76±2.12\]×106, P<0.01) in 6, 9 and 12 d cryopreservation groups after recovery for 72 h. After recovery for 24 h, the proportions of CD3+CD56+ and CD3+CD8+ cells increased gradually in all the three 6, 9 and 12 d cryopreservation groups. Moreover, both two cell proportions in 6 d cryopreservation group after recovery for 72 h was the lowest. The cytotoxicity of CIK cells on A549 cells within 24 h of recovery in all the three cryopreservation groups was much low. However, their cytotoxicity increased considerably after 24 h. The cytotoxicity of CIK cells on A549 cells in 12 d cryopreservation group was higher than those in the 6 and 9 d cryopreservation groups. For example, the anti-tumor rate of CIK cells on A549 cells in 6, 9 and 12 d cryopreservation groups after recovery for 72 h was (\[0.81±0.09\]%, \[0.59±0.06\]% and \[0.42±0.08\]%, P<0.01). ELISA assay results showed that, the levels of IFN-γ and TNF-α secreted by CIK cells increased along the recovery time in all cryopreservation groups that had been in vitro cultured for various times. After recovery for 72 h, the level of secreted IFN-γ in 6 d cryopreservation group was higher than those of the other groups, and the level of secreted TNF-α in 12 d cryopreservation group was higher than those of the other groups.Conclusion:In vitro culture of different time can influence the bioactivity of cryopreserved CIK cells. However, the bioactivity can be restored after recovery for a short time, demonstrating that CIK cells can be cryopreserved after in vitro culture for 12 d.
Objective:To observe the effect of in vitro culture of different time on proliferation, immunophenotype, cytotoxicity and cytokine secretion of human cord blood derived cytokine induced killer (CIK) cells after cryopreservation.Methods: CIK cells were cryopreserved for 1 month after in vitro culture for 6, 9 and 12 d. The cell proliferation was detected every 24 h when they were recovered and cultured for 72 h. The immunophenotype, cytotoxicity on A549 cells and the secretion of IFN-γ and TNF-α of CIK cells after recovery were detected by flow cytometry, CCK-8 assay and ELISA assay, respectively. Results:CIK cells, in vitro culture of different time before cryopreservation still showed a superior proliferation activity after recovery, among which the proliferation of CIK cells in vitro cultured for 6 d before cryopreservation, was significantly higher than those in vitro cultured for 6 and 12 d before cryopreservation. The CIK cell counts were (\[35.90±1.67\]×106, \[18.98±2.13\]×106, and \[11.76±2.12\]×106, P<0.01) in 6, 9 and 12 d cryopreservation groups after recovery for 72 h. After recovery for 24 h, the proportions of CD3+CD56+ and CD3+CD8+ cells increased gradually in all the three 6, 9 and 12 d cryopreservation groups. Moreover, both two cell proportions in 6 d cryopreservation group after recovery for 72 h was the lowest. The cytotoxicity of CIK cells on A549 cells within 24 h of recovery in all the three cryopreservation groups was much low. However, their cytotoxicity increased considerably after 24 h. The cytotoxicity of CIK cells on A549 cells in 12 d cryopreservation group was higher than those in the 6 and 9 d cryopreservation groups. For example, the anti-tumor rate of CIK cells on A549 cells in 6, 9 and 12 d cryopreservation groups after recovery for 72 h was (\[0.81±0.09\]%, \[0.59±0.06\]% and \[0.42±0.08\]%, P<0.01). ELISA assay results showed that, the levels of IFN-γ and TNF-α secreted by CIK cells increased along the recovery time in all cryopreservation groups that had been in vitro cultured for various times. After recovery for 72 h, the level of secreted IFN-γ in 6 d cryopreservation group was higher than those of the other groups, and the level of secreted TNF-α in 12 d cryopreservation group was higher than those of the other groups.Conclusion:In vitro culture of different time can influence the bioactivity of cryopreserved CIK cells. However, the bioactivity can be restored after recovery for a short time, demonstrating that CIK cells can be cryopreserved after in vitro culture for 12 d.
2013, 20(3):348-351.
DOI: 10.3872/j.issn.1007-385X.2013.03.016
Abstract:
目的： 构建靶向人角蛋白18（cytokeratin 18， CK18 ）基因的shRNA真核表达载体，稳定转染人乳腺癌MCF-7细胞株，观察其对MCF-7细胞增殖的影响。 方法： 设计两条靶向 CK18基因 的RNA干扰序列，命名为CK18-shRNA1、CK18-shRNA2，同时设计阴性对照，将合成的寡核苷酸链与Psilencer3.1-H1/Hygro质粒连接形成重组质粒，并经脂质体介导稳定转染MCF-7细胞，G418筛选后扩增获得稳定转染细胞株，分别采用RT-PCR和Western blotting法检测细胞株中 CK18 mRNA和蛋白的表达，并进一步通过MTT法检测重组表达载体稳定转染对细胞增殖的影响。 结果： 重组表达载体（Psilencer3.1-CK18-shRNA1、Psilencer3.1-CK18-shRNA2和Psilencer3.1-NC-shRNA）经PCR及DNA测序分析证明序列插入正确，经500 μg/ml G418筛选出稳定转染Psilencer3.1-CK18-shRNA的MCF-7细胞，Psilencer3.1-CK18-shRNA2转染可有效抑制MCF-7细胞中 CK18 mRNA和蛋白的表达，而Psilencer3.1-CK18-shRNA1转染不影响MCF-7细胞中 CK18 mRNA和蛋白的表达水平。与阴性对照Psilencer3.1-NC-shRNA组相比，Psilencer3.1-CK18-shRNA2转染可有效抑制MCF-7细胞的增殖\[（0.40±0.01） vs （0.55±0.06），P<0.05\]。 结论： 靶向 CK18 的shRNA表达载体稳定转染细胞后可抑制人乳腺癌MCF-7细胞的增殖。
目的： 构建靶向人角蛋白18（cytokeratin 18， CK18 ）基因的shRNA真核表达载体，稳定转染人乳腺癌MCF-7细胞株，观察其对MCF-7细胞增殖的影响。 方法： 设计两条靶向 CK18基因 的RNA干扰序列，命名为CK18-shRNA1、CK18-shRNA2，同时设计阴性对照，将合成的寡核苷酸链与Psilencer3.1-H1/Hygro质粒连接形成重组质粒，并经脂质体介导稳定转染MCF-7细胞，G418筛选后扩增获得稳定转染细胞株，分别采用RT-PCR和Western blotting法检测细胞株中 CK18 mRNA和蛋白的表达，并进一步通过MTT法检测重组表达载体稳定转染对细胞增殖的影响。 结果： 重组表达载体（Psilencer3.1-CK18-shRNA1、Psilencer3.1-CK18-shRNA2和Psilencer3.1-NC-shRNA）经PCR及DNA测序分析证明序列插入正确，经500 μg/ml G418筛选出稳定转染Psilencer3.1-CK18-shRNA的MCF-7细胞，Psilencer3.1-CK18-shRNA2转染可有效抑制MCF-7细胞中 CK18 mRNA和蛋白的表达，而Psilencer3.1-CK18-shRNA1转染不影响MCF-7细胞中 CK18 mRNA和蛋白的表达水平。与阴性对照Psilencer3.1-NC-shRNA组相比，Psilencer3.1-CK18-shRNA2转染可有效抑制MCF-7细胞的增殖\[（0.40±0.01） vs （0.55±0.06），P<0.05\]。 结论： 靶向 CK18 的shRNA表达载体稳定转染细胞后可抑制人乳腺癌MCF-7细胞的增殖。
2013, 20(3):352-355.
DOI: 10.3872/j.issn.1007-385X.2013.03.017
Abstract:
目的： 探讨人脑胶质瘤组织中胞质多聚腺苷酸化成分结合蛋白4（cytoplasmic polyadenylation element-binding protein 4，CPEB4）和B细胞淋巴瘤-XL（B-cell lymphoma-extra large，Bcl-xl）蛋白的表达及其临床意义。 方法： 收集郑州大学第五附属医院2010年1月至2012年3月手术切除且经病理证实的65例人脑胶质瘤存档蜡块（WHO Ⅰ级8例、Ⅱ级19例、Ⅲ级21例、Ⅳ级17例）和10例正常脑组织，采用免疫组织化学SP法检测CPEB4和Bcl-xl蛋白的表达，分析CPEB4和Bcl-xl与脑胶质瘤临床病理特征间的相关性。 结果： 人脑胶质瘤组织中CPEB4和Bcl-xl蛋白的阳性表达率均显著高于正常脑组织（6310% vs 000%，70.77% vs 20.00%；均P<0.01）。CPEB4和Bcl-xl蛋白的阳性表达率与患者的年龄、性别及家族史均无明显相关性（P>0.05），但与肿瘤的恶性程度呈正相关（P<0.05）。人脑胶质瘤组织中CPEB4和Bcl-xl蛋白的表达呈正相关关系（r=0419，P=0.001）。 结论： CPEB4和Bcl-xl在人脑胶质瘤组织中的阳性表达率明显增高，且与肿瘤恶性程度呈正相关，提示CPEB4可能与Bcl-xl蛋白相互作用，在人脑胶质瘤的发生、发展中起重要作用。
目的： 探讨人脑胶质瘤组织中胞质多聚腺苷酸化成分结合蛋白4（cytoplasmic polyadenylation element-binding protein 4，CPEB4）和B细胞淋巴瘤-XL（B-cell lymphoma-extra large，Bcl-xl）蛋白的表达及其临床意义。 方法： 收集郑州大学第五附属医院2010年1月至2012年3月手术切除且经病理证实的65例人脑胶质瘤存档蜡块（WHO Ⅰ级8例、Ⅱ级19例、Ⅲ级21例、Ⅳ级17例）和10例正常脑组织，采用免疫组织化学SP法检测CPEB4和Bcl-xl蛋白的表达，分析CPEB4和Bcl-xl与脑胶质瘤临床病理特征间的相关性。 结果： 人脑胶质瘤组织中CPEB4和Bcl-xl蛋白的阳性表达率均显著高于正常脑组织（6310% vs 000%，70.77% vs 20.00%；均P<0.01）。CPEB4和Bcl-xl蛋白的阳性表达率与患者的年龄、性别及家族史均无明显相关性（P>0.05），但与肿瘤的恶性程度呈正相关（P<0.05）。人脑胶质瘤组织中CPEB4和Bcl-xl蛋白的表达呈正相关关系（r=0419，P=0.001）。 结论： CPEB4和Bcl-xl在人脑胶质瘤组织中的阳性表达率明显增高，且与肿瘤恶性程度呈正相关，提示CPEB4可能与Bcl-xl蛋白相互作用，在人脑胶质瘤的发生、发展中起重要作用。
2013, 20(3):356-359.
DOI: 10.3872/j.issn.1007-385X.2013.03.018
Abstract:
核因子κB（nuclear factor- κB, NF-κB）是一种重要的核转录因子，在调控细胞周期、凋亡和代谢等生理过程中发挥重要作用；然而，NF-κB在人类多种恶性肿瘤中过表达，在肿瘤的发生、发展和免疫炎症反应中也起着重要的调控作用。在结直肠癌中NF-κB同样呈高表达，且呈激活状态。NF-κB信号通路被激活后，NF-κB转入细胞核内，通过与下游靶基因结合，促进目的基因的转录，从而调控目的蛋白的表达，并介导多条信号通路，促进肿瘤细胞血管生成、增加肿瘤细胞增殖、抗凋亡和促进肿瘤细胞的侵袭、迁移和转移能力，以及对放化疗的抵抗，甚至诱导肿瘤细胞获得干性等恶性表型。研究表明，NF-κB过表达的结直肠癌患者病情进展较快，生存时间较短，NF-κB可作为结直肠癌患者疗效监测和预后监测的一个生物标志物，针对NF-κB及其信号通路的分子靶向治疗为结直肠癌患者治疗提供潜在的策略。
核因子κB（nuclear factor- κB, NF-κB）是一种重要的核转录因子，在调控细胞周期、凋亡和代谢等生理过程中发挥重要作用；然而，NF-κB在人类多种恶性肿瘤中过表达，在肿瘤的发生、发展和免疫炎症反应中也起着重要的调控作用。在结直肠癌中NF-κB同样呈高表达，且呈激活状态。NF-κB信号通路被激活后，NF-κB转入细胞核内，通过与下游靶基因结合，促进目的基因的转录，从而调控目的蛋白的表达，并介导多条信号通路，促进肿瘤细胞血管生成、增加肿瘤细胞增殖、抗凋亡和促进肿瘤细胞的侵袭、迁移和转移能力，以及对放化疗的抵抗，甚至诱导肿瘤细胞获得干性等恶性表型。研究表明，NF-κB过表达的结直肠癌患者病情进展较快，生存时间较短，NF-κB可作为结直肠癌患者疗效监测和预后监测的一个生物标志物，针对NF-κB及其信号通路的分子靶向治疗为结直肠癌患者治疗提供潜在的策略。
2013, 20(3):360-364.
DOI: 10.3872/j.issn.1007-385X.2013.03.019
Abstract:
贫血是多发性骨髓瘤（multiple myeloma，MM）的常见症状，与预后不良相关。目前认为，MM相关性贫血是一种慢性病性贫血（anemia of chronic disease，ACD），由慢性炎症刺激所导致。铁调素（hepcidin）是一种机体铁代谢调节激素，它在炎症刺激下表达上调，使铁无法从细胞释放至血浆，造成血清铁降低，进而导致贫血发生。铁调素相关通路是MM相关性贫血发病机制中的中心环节，成为MM相关性贫血治疗的新靶点。促红细胞生成素类制剂（erythropoiesis-stimulating agent，ESA）能有效提高血红蛋白水平，是既往治疗MM相关性贫血的主要方法之一。近年ESA对肿瘤相关性贫血的治疗效果受到质疑，认为ESA可能促进肿瘤进展，并不能改善患者的总体预后。本文主要对MM相关性贫血的发病机制及治疗进行综述，着重阐述铁调素的作用机制，总结ESA的治疗效果，探讨铁调素通路的靶向治疗作用。
贫血是多发性骨髓瘤（multiple myeloma，MM）的常见症状，与预后不良相关。目前认为，MM相关性贫血是一种慢性病性贫血（anemia of chronic disease，ACD），由慢性炎症刺激所导致。铁调素（hepcidin）是一种机体铁代谢调节激素，它在炎症刺激下表达上调，使铁无法从细胞释放至血浆，造成血清铁降低，进而导致贫血发生。铁调素相关通路是MM相关性贫血发病机制中的中心环节，成为MM相关性贫血治疗的新靶点。促红细胞生成素类制剂（erythropoiesis-stimulating agent，ESA）能有效提高血红蛋白水平，是既往治疗MM相关性贫血的主要方法之一。近年ESA对肿瘤相关性贫血的治疗效果受到质疑，认为ESA可能促进肿瘤进展，并不能改善患者的总体预后。本文主要对MM相关性贫血的发病机制及治疗进行综述，着重阐述铁调素的作用机制，总结ESA的治疗效果，探讨铁调素通路的靶向治疗作用。
2013, 20(3):365-371.
DOI: 10.3872/j.issn.1007-385X.2013.03.020
Abstract:
黑素瘤是一种侵袭性强的难治性皮肤肿瘤，其自然病程中出现的与淋巴细胞浸润相关的肿瘤自发回缩或消退现象，以及黑素瘤对各种免疫治疗较好的临床反应均提示了黑素瘤是一种免疫原性肿瘤。本文从主动免疫治疗和被动免疫治疗两个方面综述了近年来黑素瘤的免疫治疗进展。在以T细胞过继免疫和疫苗为中心的肿瘤抗原特异性主动免疫治疗方法中，肿瘤浸润淋巴细胞（tumor infiltrating lymphocyte，TIL）体外扩增是一种较为稳定且有效的方式，但存在TIL不易获取、培养不易成功等局限性；DC疫苗因为能够诱导并促进肿瘤特异性CTL扩增而被视作是一种有前途的治疗方式，但仍然面临临床疗效不够满意等问题，随着新的特异性抗原的发现及抗原负载方式的改进，相信DC疫苗将会使患者更多获益。而新的特异性抗原的发现，也必将推动T细胞抗原受体（T cell antigen receptor，TCR）基因修饰的T细胞和疫苗的研发进展；调节分子IL-21和针对细胞毒性T细胞抗原-4（cytotoxic T lymphocyte-associated antigen-4，CTLA-4）、程序性死亡分子1（programmed cell death-1，PD-1）、CD40的单克隆抗体研究显示出一定的临床潜力，也为黑素瘤的免疫治疗打开了新的思路。
黑素瘤是一种侵袭性强的难治性皮肤肿瘤，其自然病程中出现的与淋巴细胞浸润相关的肿瘤自发回缩或消退现象，以及黑素瘤对各种免疫治疗较好的临床反应均提示了黑素瘤是一种免疫原性肿瘤。本文从主动免疫治疗和被动免疫治疗两个方面综述了近年来黑素瘤的免疫治疗进展。在以T细胞过继免疫和疫苗为中心的肿瘤抗原特异性主动免疫治疗方法中，肿瘤浸润淋巴细胞（tumor infiltrating lymphocyte，TIL）体外扩增是一种较为稳定且有效的方式，但存在TIL不易获取、培养不易成功等局限性；DC疫苗因为能够诱导并促进肿瘤特异性CTL扩增而被视作是一种有前途的治疗方式，但仍然面临临床疗效不够满意等问题，随着新的特异性抗原的发现及抗原负载方式的改进，相信DC疫苗将会使患者更多获益。而新的特异性抗原的发现，也必将推动T细胞抗原受体（T cell antigen receptor，TCR）基因修饰的T细胞和疫苗的研发进展；调节分子IL-21和针对细胞毒性T细胞抗原-4（cytotoxic T lymphocyte-associated antigen-4，CTLA-4）、程序性死亡分子1（programmed cell death-1，PD-1）、CD40的单克隆抗体研究显示出一定的临床潜力，也为黑素瘤的免疫治疗打开了新的思路。
2013, 20(3):372-375.
DOI: 10.3872/j.issn.1007-385X.2013.03.021
Abstract:
淋巴细胞归巢受机体组织微环境的调控，肿瘤形成时机体微环境的变化影响淋巴细胞的迁移和浸润。肿瘤浸润淋巴细胞（tumor-infiltration lymphocyte，TIL）作为重要的抗肿瘤免疫细胞，其分布和浸润程度与肿瘤发生、发展以及预后密切相关。淋巴细胞浸润肿瘤的调节机制可能涉及肿瘤脉管对淋巴细胞浸润的正反两方面的调节，黏附分子和趋化因子介导淋巴细胞外渗、募集、肿瘤抗原定位淋巴细胞浸润程度以及其他免疫细胞影响淋巴细胞浸润等。如何引导淋巴细胞向肿瘤迁移、能否创造利于瘤内浸润的微环境等，对于加速TIL临床转化，使肿瘤患者更大程度获益具有重要意义。
淋巴细胞归巢受机体组织微环境的调控，肿瘤形成时机体微环境的变化影响淋巴细胞的迁移和浸润。肿瘤浸润淋巴细胞（tumor-infiltration lymphocyte，TIL）作为重要的抗肿瘤免疫细胞，其分布和浸润程度与肿瘤发生、发展以及预后密切相关。淋巴细胞浸润肿瘤的调节机制可能涉及肿瘤脉管对淋巴细胞浸润的正反两方面的调节，黏附分子和趋化因子介导淋巴细胞外渗、募集、肿瘤抗原定位淋巴细胞浸润程度以及其他免疫细胞影响淋巴细胞浸润等。如何引导淋巴细胞向肿瘤迁移、能否创造利于瘤内浸润的微环境等，对于加速TIL临床转化，使肿瘤患者更大程度获益具有重要意义。
2013, 20(3):376-380.
DOI: 10.3872/j.issn.1007-385X.2013.03.022
Abstract:
磷脂酰乙醇胺结合蛋白1（phosphatidylethanolamine-binding protein，PEBP1）广泛表达于多种生物中，具有重要的生理学功能。PEBP1可以与Raf-1结合，从而抑制MAPK信号转导通路，并参与对G蛋白偶联受体、 NF-κB和GSK3β 等多条信号通路的调控。近年来的研究发现，PEBP1在肿瘤中发挥重要作用，PEBP1通过TNF-α，FasL（Fas ligand）和TNF相关的凋亡诱导配体（TNF-related apoptosis-inducing ligand，TRAIL）等死亡配体促进肿瘤细胞的凋亡，同时PEBP1作为肿瘤转移的抑制基因，在肿瘤组织中表达较正常组织明显减少，因而PEBP1也成为一个新的肿瘤标志物。此外，PEBP1通过调控 MAPK和NF-κB 等信号通路，影响细胞的分化以及细胞分裂和基因组稳定性。本文就PEBP1在肿瘤研究中的一些最新研究进展作一综述。
磷脂酰乙醇胺结合蛋白1（phosphatidylethanolamine-binding protein，PEBP1）广泛表达于多种生物中，具有重要的生理学功能。PEBP1可以与Raf-1结合，从而抑制MAPK信号转导通路，并参与对G蛋白偶联受体、 NF-κB和GSK3β 等多条信号通路的调控。近年来的研究发现，PEBP1在肿瘤中发挥重要作用，PEBP1通过TNF-α，FasL（Fas ligand）和TNF相关的凋亡诱导配体（TNF-related apoptosis-inducing ligand，TRAIL）等死亡配体促进肿瘤细胞的凋亡，同时PEBP1作为肿瘤转移的抑制基因，在肿瘤组织中表达较正常组织明显减少，因而PEBP1也成为一个新的肿瘤标志物。此外，PEBP1通过调控 MAPK和NF-κB 等信号通路，影响细胞的分化以及细胞分裂和基因组稳定性。本文就PEBP1在肿瘤研究中的一些最新研究进展作一综述。
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
- 邮编：200433
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
您是第位访问者
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.