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Volume 26,Issue 8,2019 Table of Contents
2019, 26(8):833-836.
DOI: 10.3872/j.issn.1007-385X.2019.08.001
Abstract:
[Abstract] Chimeric antigen receptor (CAR) T lymphocyte has shown attractive prospects in the treatment of lymphohematopoietic malignancies including B-cell lymphoblastic leukemia, B-cell lymphoma and multiple myeloma. Many applicants have submitted investigational new drug (IND) applications to Center for Drug Evaluation of National Medical Products Ggency, however, many of the INDs have problems in patient selection, prognostic indicators and risk management, etc, which might hinder the evaluation of the safety and efficacy of CAR-T cells. Thus, we made some suggestions on the above-mentioned problems through summarizing clinical experience and communicating with domestic clinical experts, which the sponsors and researchers can refer to when conducting CAR-T cell clinical trials for registration.
[Abstract] Chimeric antigen receptor (CAR) T lymphocyte has shown attractive prospects in the treatment of lymphohematopoietic malignancies including B-cell lymphoblastic leukemia, B-cell lymphoma and multiple myeloma. Many applicants have submitted investigational new drug (IND) applications to Center for Drug Evaluation of National Medical Products Ggency, however, many of the INDs have problems in patient selection, prognostic indicators and risk management, etc, which might hinder the evaluation of the safety and efficacy of CAR-T cells. Thus, we made some suggestions on the above-mentioned problems through summarizing clinical experience and communicating with domestic clinical experts, which the sponsors and researchers can refer to when conducting CAR-T cell clinical trials for registration.
2019, 26(8):837-844.
DOI: 10.3872/j.issn.1007-385X.2019.08.002
Abstract:
[Abstract] Objective ：To investigate the role and mechanism of chromosomal region maintenance 1 (CRM1) inhibitor sulforaphene (LFS-01) in killing triple negative breast cancer (TNBC) cells by inhibiting signal transducer and activator of transcription 3 (STAT3) signaling pathways. Methods: Whether LFS-01 could combine with the NES pocket of CRM1 was verified by molecular dynamics simulation techniques. The killing activity of LFS-01 on four different breast cancer cell lines was detected by CCK-8 method. TNBC MDA-MB-468 and MDA-MB-231 cells were treated with different concentrations of LFS-01, and the intracellular localization of CRM1 cargo protein STAT3 and protein with NES sequence was detected by immunofluorescence; WB was used to detect the effect of LFS-01 on the expression of STAT-3 signaling pathway and its downstream proteins; WB, cellular immunofluorescence and transmission electron microscopy were adopted to detect the occurrence of autophagy; the effect of LFS-01 on cell cycle and apoptosis was detected by flow cytometry. Results: Molecular dynamics simulations showed that LFS-01 can bind to the NES pocket of CRM1, indicating that it may structurally affect the latter's protein transport function. LFS-01 could specifically kill TNBC MDA-MB-468 and MDA-MB-231 cells. STAT3 and NES-tagged proteins were mainly blocked in the nucleus of TNBC cells after the treatment with 10 μmol/L LFS-01, while they were evenly distributed in the cytoplasm in the control group. The expressions of phosphorylated STAT3 protein, Bcl-xL and Cylin D1 were decreased in MDA-MB-468 and MDA-MB-231 cells with the increase of LFS-01 dose and the prolongation of treatment time; the expression of autophagy marker protein LC3B increased, and high-density, multi-layered autophagosomes appeared at the same time; cell cycle arrest was observed in S phase and apoptosis rate was significantly increased (P<0.05 or P<0.01). Conclusion: LFS-01 blocks the export of CRM1 carrier protein, thereby inhibiting the activation of STAT3 signaling pathway and promoting autophagy, cell cycle arrest and apoptosis in TNBC MDA-MB-468 and MDAMB-231 cells.
[Abstract] Objective ：To investigate the role and mechanism of chromosomal region maintenance 1 (CRM1) inhibitor sulforaphene (LFS-01) in killing triple negative breast cancer (TNBC) cells by inhibiting signal transducer and activator of transcription 3 (STAT3) signaling pathways. Methods: Whether LFS-01 could combine with the NES pocket of CRM1 was verified by molecular dynamics simulation techniques. The killing activity of LFS-01 on four different breast cancer cell lines was detected by CCK-8 method. TNBC MDA-MB-468 and MDA-MB-231 cells were treated with different concentrations of LFS-01, and the intracellular localization of CRM1 cargo protein STAT3 and protein with NES sequence was detected by immunofluorescence; WB was used to detect the effect of LFS-01 on the expression of STAT-3 signaling pathway and its downstream proteins; WB, cellular immunofluorescence and transmission electron microscopy were adopted to detect the occurrence of autophagy; the effect of LFS-01 on cell cycle and apoptosis was detected by flow cytometry. Results: Molecular dynamics simulations showed that LFS-01 can bind to the NES pocket of CRM1, indicating that it may structurally affect the latter's protein transport function. LFS-01 could specifically kill TNBC MDA-MB-468 and MDA-MB-231 cells. STAT3 and NES-tagged proteins were mainly blocked in the nucleus of TNBC cells after the treatment with 10 μmol/L LFS-01, while they were evenly distributed in the cytoplasm in the control group. The expressions of phosphorylated STAT3 protein, Bcl-xL and Cylin D1 were decreased in MDA-MB-468 and MDA-MB-231 cells with the increase of LFS-01 dose and the prolongation of treatment time; the expression of autophagy marker protein LC3B increased, and high-density, multi-layered autophagosomes appeared at the same time; cell cycle arrest was observed in S phase and apoptosis rate was significantly increased (P<0.05 or P<0.01). Conclusion: LFS-01 blocks the export of CRM1 carrier protein, thereby inhibiting the activation of STAT3 signaling pathway and promoting autophagy, cell cycle arrest and apoptosis in TNBC MDA-MB-468 and MDAMB-231 cells.
2019, 26(8):845-849.
DOI: 10.3872/j.issn.1007-385X.2019.08.003
Abstract:
[Abstract] Objective: To investigate the effects of long non-coding RNA nuclear enriched abundant transcript 1 (lncRNA NEAT1) on the proliferation of lung adenocarcinoma PC-9 cells and to explore its mechanism. Methods: qPCR was used to detect the expression level of lncRNA NEAT1 in human lung adenocarcinoma PC-9 cells and human embryonic lung diploid 2BS cells. The sequence of small interfering RNA (siRNA) targeting lncRNA NEAT1 gene was designed and synthesized, and then transfected into PC-9 cells by liposome method. The expression level of NEAT1 in PC-9 cells before and after transfection was detected by qPCR. MTT and flow cytometry were used to detect the effect of lncRNA NEAT1 knockdown on proliferation and cell cycle distribution of PC-9 cells, respectively.WB assay was used to detect the expressions of DNA damage-related proteins, namely, double-stranded DNA breaks (DSBs)biomarker γ -H2AX and ataxia-telangiectasia mutated (ATM), before and after transfection. Results: Compared with 2BS cells, lncRNA NEAT1 was highly expressed in PC-9 cells (P<0.05). The PC-9 cells with lncRNA NEAT1 knock-down were successfully established. After being transfected with siRNA for 12 h, the proliferation of PC-9 cells in siNEAT1 group and siNEAT2 group significantly decreased as compared with the blank control group and the empty transfection group (P<0.05). In the interference groups,cell cycle was arrested in G1 phase ([88.97±2.64]%, [88.15±1.48]% vs [84.5±1.72]%, P<0.05) and G2/M phase ([8.35±2.02]%, [8.11±1.36]% vs [4.28±1.28]%, P<0.05). The expression levels of DNA damage-related proteins ATM and γ-H2AX in the interference groups were significantly increased (all P<0.05). Conclusion: lncRNA NEAT1 is highly expressed in lung adenocarcinoma PC-9 cells. lncRNA NEAT1 inhibits DNA damage and causes cell cycle at G1/M phase switch, and thus promotes the proliferation of lung adenocarcinoma cells.
[Abstract] Objective: To investigate the effects of long non-coding RNA nuclear enriched abundant transcript 1 (lncRNA NEAT1) on the proliferation of lung adenocarcinoma PC-9 cells and to explore its mechanism. Methods: qPCR was used to detect the expression level of lncRNA NEAT1 in human lung adenocarcinoma PC-9 cells and human embryonic lung diploid 2BS cells. The sequence of small interfering RNA (siRNA) targeting lncRNA NEAT1 gene was designed and synthesized, and then transfected into PC-9 cells by liposome method. The expression level of NEAT1 in PC-9 cells before and after transfection was detected by qPCR. MTT and flow cytometry were used to detect the effect of lncRNA NEAT1 knockdown on proliferation and cell cycle distribution of PC-9 cells, respectively.WB assay was used to detect the expressions of DNA damage-related proteins, namely, double-stranded DNA breaks (DSBs)biomarker γ -H2AX and ataxia-telangiectasia mutated (ATM), before and after transfection. Results: Compared with 2BS cells, lncRNA NEAT1 was highly expressed in PC-9 cells (P<0.05). The PC-9 cells with lncRNA NEAT1 knock-down were successfully established. After being transfected with siRNA for 12 h, the proliferation of PC-9 cells in siNEAT1 group and siNEAT2 group significantly decreased as compared with the blank control group and the empty transfection group (P<0.05). In the interference groups,cell cycle was arrested in G1 phase ([88.97±2.64]%, [88.15±1.48]% vs [84.5±1.72]%, P<0.05) and G2/M phase ([8.35±2.02]%, [8.11±1.36]% vs [4.28±1.28]%, P<0.05). The expression levels of DNA damage-related proteins ATM and γ-H2AX in the interference groups were significantly increased (all P<0.05). Conclusion: lncRNA NEAT1 is highly expressed in lung adenocarcinoma PC-9 cells. lncRNA NEAT1 inhibits DNA damage and causes cell cycle at G1/M phase switch, and thus promotes the proliferation of lung adenocarcinoma cells.
2019, 26(8):850-855.
DOI: 10.3872/j.issn.1007-385X.2019.08.004
Abstract:
[Abstract] Objective: To investigate the effects of GBX2 gene on the proliferation, migration and invasion of human cervical carcinoma SiHa cells and to explore the mechanism. Methods: Recombinant plasmid over-expressing GBX2 gene (pCMV6-entry-GBX2,experimental group) and empty vector plasmid (pCMV6-entry, negative control group) were transfected into cervical cancer SiHa cells by plasmid transfection technique. The proliferation, colony formation and cell cycle of transfected cells were detected by WST-1 method,Colony formation assay and flow cytometry, respectively. The cell migration and invasion were detected by wound healing assay and Transwell assay. The expression level of IL-6 in cell culture supernatant was detected by ELISA. WB was used to detect the expression changes of EMT-related proteins and to explore its possible mechanism. Results: Compared with the SiHa/pCMV6 negative control group, after up-regulation of GBX2, (1) the proliferation, colony formation, migration and invasion of SiHa/GBX2 cells in the experimental group were significantly enhanced (all P<0.01); The proportion of cells in G0/G1 phase decreased while the proportion of cells in S phase and G2/M phase increased (all P<0.01); (2) the expression of E-cadherin decreased, and the expressions of N-cadherin, vimentin and snail increased (all P<0.01); (3) the expression of IL-6 in the culture supernatant of SiHa/GBX2 cells was significantly up-regulated (P<0.01); (4) STAT3 phosphorylation in SiHa/GBX2 cells was enhanced, and could be inhibited by STAT3 inhibitor S31-201 (P<0.01). Conclusion: GBX2 may induce EMT of cervical cancer SiHa cells through IL-6/STAT3 pathway, thereby promoting the proliferation, migration and invasion of cervical cancer cells.
[Abstract] Objective: To investigate the effects of GBX2 gene on the proliferation, migration and invasion of human cervical carcinoma SiHa cells and to explore the mechanism. Methods: Recombinant plasmid over-expressing GBX2 gene (pCMV6-entry-GBX2,experimental group) and empty vector plasmid (pCMV6-entry, negative control group) were transfected into cervical cancer SiHa cells by plasmid transfection technique. The proliferation, colony formation and cell cycle of transfected cells were detected by WST-1 method,Colony formation assay and flow cytometry, respectively. The cell migration and invasion were detected by wound healing assay and Transwell assay. The expression level of IL-6 in cell culture supernatant was detected by ELISA. WB was used to detect the expression changes of EMT-related proteins and to explore its possible mechanism. Results: Compared with the SiHa/pCMV6 negative control group, after up-regulation of GBX2, (1) the proliferation, colony formation, migration and invasion of SiHa/GBX2 cells in the experimental group were significantly enhanced (all P<0.01); The proportion of cells in G0/G1 phase decreased while the proportion of cells in S phase and G2/M phase increased (all P<0.01); (2) the expression of E-cadherin decreased, and the expressions of N-cadherin, vimentin and snail increased (all P<0.01); (3) the expression of IL-6 in the culture supernatant of SiHa/GBX2 cells was significantly up-regulated (P<0.01); (4) STAT3 phosphorylation in SiHa/GBX2 cells was enhanced, and could be inhibited by STAT3 inhibitor S31-201 (P<0.01). Conclusion: GBX2 may induce EMT of cervical cancer SiHa cells through IL-6/STAT3 pathway, thereby promoting the proliferation, migration and invasion of cervical cancer cells.
2019, 26(8):856-861.
DOI: 10.3872/j.issn.1007-385X.2019.08.005
Abstract:
[Abstract] Objective: To investigate the effect of enolase 1 (ENO1) expression on proliferation, apoptosis and migration of lung cancer PC14 cells. Methods: ENO1 over-expression vector-pcDNA3.1/ENO1 was constructed and transfected into PC14 cells at logarithmic growth phase with liposome LipofectamineTM 2000. G418 was used to screen PC14 cells that stably expressing ENO1. The effects of ENO1 over-expression on proliferation, migration and apoptosis of PC14 cells were detected by CCK-8 method, scratch-healing assay and flow cytometry, respectively. Results: The ENO1 over-expression cell model was successfully constructed. Compared with PC14-vehicle and wild-type PC14 cells, the mRNA and protein expression levels of ENO1 in PC14-ENO1 cells were significantly elevated (all P<0.05), and the proliferation of PC14-ENO1 cells was significantly increased (all P<0.05). The relative mobility of PC14-ENO1 cells was significantly higher than that of pcDNA3.1-vehicle cells and wild-type PC14 cells ([13.26±1.13]% vs [8.46±1.11]%,[7.86±1.00]%, both P<0.05). There was no significant difference in apoptotic rate among PC14-ENO1, PC14-vehicle and PC14 cells (all P > 0.05) Conclusion: Over-expression of ENO1 promotes proliferation and migration of lung cancer PC14 cells.
[Abstract] Objective: To investigate the effect of enolase 1 (ENO1) expression on proliferation, apoptosis and migration of lung cancer PC14 cells. Methods: ENO1 over-expression vector-pcDNA3.1/ENO1 was constructed and transfected into PC14 cells at logarithmic growth phase with liposome LipofectamineTM 2000. G418 was used to screen PC14 cells that stably expressing ENO1. The effects of ENO1 over-expression on proliferation, migration and apoptosis of PC14 cells were detected by CCK-8 method, scratch-healing assay and flow cytometry, respectively. Results: The ENO1 over-expression cell model was successfully constructed. Compared with PC14-vehicle and wild-type PC14 cells, the mRNA and protein expression levels of ENO1 in PC14-ENO1 cells were significantly elevated (all P<0.05), and the proliferation of PC14-ENO1 cells was significantly increased (all P<0.05). The relative mobility of PC14-ENO1 cells was significantly higher than that of pcDNA3.1-vehicle cells and wild-type PC14 cells ([13.26±1.13]% vs [8.46±1.11]%,[7.86±1.00]%, both P<0.05). There was no significant difference in apoptotic rate among PC14-ENO1, PC14-vehicle and PC14 cells (all P > 0.05) Conclusion: Over-expression of ENO1 promotes proliferation and migration of lung cancer PC14 cells.
2019, 26(8):862-867.
DOI: 10.3872/j.issn.1007-385X.2019.08.006
Abstract:
[Abstract] Objective: To investigate the role and mechanism of Krüppel-like factor 4 (KLF4) in regulating epithelial-mesenchymal transition (EMT) and migration of bladder cancer cells. Methods: Bladder cancer 5637 and T24 cell lines that stably over-expressing KLF4 (LV-KLF4, experiment group) were constructed, and the negative control group (LV-NC) was also established; the mRNA and protein expressions of KLF4 were verified by qPCR and WB, respectively. Transwell chamber assay was used to detect the migration ability of cells in LV-KLF4 and LV-NC groups. WB was performed to detect the expression levels of EMT-related markers (E-cadherin, N-cadherin, Vimentin) and Wnt signaling pathway-related proteins. Immunofluorescence technique was used to detect the distribution of β-catenin in cells after over-expression of KLF4. Results: The 5637 and T24 cell lines over-expressing KLF4 gene were successfully constructed. Compared with the LV-NC group, the mRNA and protein expressions of KLF4 increased in LV-KLF4 groups (all P<0.01); the expression of E-cadherin increased (P<0.01), while the expressions of N-cadherin, vimentin, and the expression levels of total β - catenin, nuclear β - catenin, MMP 9 and c-Myc decreased (all P<0.01); moreover, the migration ability of cells decreased significantly (P<0.01); the fluorescence expression of β -catenin in cells also decreased significantly in LV-KLF4 group as compared to LV-NC group. Conclusion: Over-expression of KLF4 gene in bladder cancer cells may inhibit EMT process by regulating Wnt/ β -catenin signaling pathway, and further inhibit the migration of bladder cancer 5637 and T24 cells.
[Abstract] Objective: To investigate the role and mechanism of Krüppel-like factor 4 (KLF4) in regulating epithelial-mesenchymal transition (EMT) and migration of bladder cancer cells. Methods: Bladder cancer 5637 and T24 cell lines that stably over-expressing KLF4 (LV-KLF4, experiment group) were constructed, and the negative control group (LV-NC) was also established; the mRNA and protein expressions of KLF4 were verified by qPCR and WB, respectively. Transwell chamber assay was used to detect the migration ability of cells in LV-KLF4 and LV-NC groups. WB was performed to detect the expression levels of EMT-related markers (E-cadherin, N-cadherin, Vimentin) and Wnt signaling pathway-related proteins. Immunofluorescence technique was used to detect the distribution of β-catenin in cells after over-expression of KLF4. Results: The 5637 and T24 cell lines over-expressing KLF4 gene were successfully constructed. Compared with the LV-NC group, the mRNA and protein expressions of KLF4 increased in LV-KLF4 groups (all P<0.01); the expression of E-cadherin increased (P<0.01), while the expressions of N-cadherin, vimentin, and the expression levels of total β - catenin, nuclear β - catenin, MMP 9 and c-Myc decreased (all P<0.01); moreover, the migration ability of cells decreased significantly (P<0.01); the fluorescence expression of β -catenin in cells also decreased significantly in LV-KLF4 group as compared to LV-NC group. Conclusion: Over-expression of KLF4 gene in bladder cancer cells may inhibit EMT process by regulating Wnt/ β -catenin signaling pathway, and further inhibit the migration of bladder cancer 5637 and T24 cells.
2019, 26(8):868-875.
DOI: 10.3872/j.issn.1007-385X.2019.08.007
Abstract:
[Abstract] Objective: To explore the role of tumor suppressor gene programmed cell death 5 gene (PDCD5) in the growth and temozolomide (TMZ) sensitivity of brain glioma cells. Methods: A total of 116 patients with cerebral glioma admitted to the Department of Neurosurgery, First Clinical Hospital of Jilin University from January 2009 to December 2014 were enrolled in this study. QPCR, WB and immunohistochemistry method were used to detect the mRNA and protein expressions of PDCD5 in glioma cell lines (U87, U251),U87 cell line with stable PDCD5 expression (U87-PDCD5), glioma cells with si-PDCD5 transfection and primary cerebral glioma tissues,respectively. MTT assay was used to detect the effect of over-expression or knockdown of PDCD5 on the growth and TMZ-sensitivity of glioma cells. The subcutaneous tumor-bearing model of glioma cell line U87 was established in nude mice, and then the experimental mice were randomly divided into control group, TMZ group, PDCD5 group and TMZ+exogenous PDCD5 recombinant expres-sion vector group. After 20 days, the animals were sacrificed by cervical dislocation and the tumor tissue was excised to measure the tumor volume and weigh. The expression of PDCD5 in tumor tissues was detected by qPCR and WB methods, and the effects of PDCD5 combined with TMZ on the growth of gliomas were also analyzed. Results: The relative mRNA and protein expressions of PDCD5 in U87 cells were significantly lower than those in U251 cells (both P<0.05), and the mRNA and protein expressions of PDCD5 in high level glioma tissues were significantly lower than those in low level tissues (all P<0.05). The sensitivity of U87-PDCD5 cells and U251 cells to TMZ was higher than that of U87 cells (all P<0.05). The sensitivity of cells to TMZ in U87-PDCD5-siRNA group and U251-siRNA group was significantly lower than that of the control group (both P<0.05). The tumor volume and weight of nude mice xenografts were compared, and the results showed control group>TMZ group>PDCD5 group>combined group (all P<0.05); however, the mRNAand protein expressions of PDCD5 in the transplanted tumor tissues of each group showed the opposite trend (all P<0.05). Conclusion:PDCD5 over-expression can enhance the chemosensitivity of brain glioma to the chemotherapy drug TMZ, while silencing of PDCD5 expression exerts the opposite effect.Thecombination of PDCD5 and TMZ can better inhibit the growth of xenografts in nude mice.
[Abstract] Objective: To explore the role of tumor suppressor gene programmed cell death 5 gene (PDCD5) in the growth and temozolomide (TMZ) sensitivity of brain glioma cells. Methods: A total of 116 patients with cerebral glioma admitted to the Department of Neurosurgery, First Clinical Hospital of Jilin University from January 2009 to December 2014 were enrolled in this study. QPCR, WB and immunohistochemistry method were used to detect the mRNA and protein expressions of PDCD5 in glioma cell lines (U87, U251),U87 cell line with stable PDCD5 expression (U87-PDCD5), glioma cells with si-PDCD5 transfection and primary cerebral glioma tissues,respectively. MTT assay was used to detect the effect of over-expression or knockdown of PDCD5 on the growth and TMZ-sensitivity of glioma cells. The subcutaneous tumor-bearing model of glioma cell line U87 was established in nude mice, and then the experimental mice were randomly divided into control group, TMZ group, PDCD5 group and TMZ+exogenous PDCD5 recombinant expres-sion vector group. After 20 days, the animals were sacrificed by cervical dislocation and the tumor tissue was excised to measure the tumor volume and weigh. The expression of PDCD5 in tumor tissues was detected by qPCR and WB methods, and the effects of PDCD5 combined with TMZ on the growth of gliomas were also analyzed. Results: The relative mRNA and protein expressions of PDCD5 in U87 cells were significantly lower than those in U251 cells (both P<0.05), and the mRNA and protein expressions of PDCD5 in high level glioma tissues were significantly lower than those in low level tissues (all P<0.05). The sensitivity of U87-PDCD5 cells and U251 cells to TMZ was higher than that of U87 cells (all P<0.05). The sensitivity of cells to TMZ in U87-PDCD5-siRNA group and U251-siRNA group was significantly lower than that of the control group (both P<0.05). The tumor volume and weight of nude mice xenografts were compared, and the results showed control group>TMZ group>PDCD5 group>combined group (all P<0.05); however, the mRNAand protein expressions of PDCD5 in the transplanted tumor tissues of each group showed the opposite trend (all P<0.05). Conclusion:PDCD5 over-expression can enhance the chemosensitivity of brain glioma to the chemotherapy drug TMZ, while silencing of PDCD5 expression exerts the opposite effect.Thecombination of PDCD5 and TMZ can better inhibit the growth of xenografts in nude mice.
2019, 26(8):876-881.
DOI: 10.3872/j.issn.1007-385X.2019.08.008
Abstract:
[Abstract] Objective：To study the regulatory effect of mogrol (MO) on lipid metabolism of hepatic cancer cells and its molecular mechanism. Methods: Oleic acid (OA) was used to induce fat accumulation in hepatocellular carcinoma HepG2 cells and to establish a steatosis cell model. CCK-8 method was used to detect the cytotoxicity of MO to HepG2 cells, and an experimental working concentration without obvious cytotoxicity of MO was chosen. After being treated with different concentrations of MO, lipid accumulation in the cells was observed by oil red O staining, and the contents of triglyceride (TG) and cholesterol (TC) in the cells were measured. Key genes involving in lipid metabolism were screened out by high-throughput transcriptome sequencing qPCR was used to detect the mRNA expressions of ,SREBP-1c and FASN, while Western Blot was used to detect the protein expressions of p-AMPKα, SREBP-1c and FASN in cells of model group and treatment group. Results: After OA induction, a large amount of lipids accumulated in HepG2 cells, the contents of TG and TC increased significantly. Three key genes (SREBP-1c, FASN and p-AMPK α) involving in lipid metabolism of hepatic cancer cells were screened out. After OA induction, the mRNA expressions of SREBP-1c and FASN increased, the protein expression of p-AMPK α decreased while the protein expressions of SREBP-1c, FASN and other proteins increased significantly.After intervention with working concentration of MO, intracellular lipid accumulation, contents of TG and TC, mRNA expressions of SREBP-1c, FASN and protein expressions of SREBP-1c, FASN decreased significantly, while the expression of p-AMPKα increased.Conclusion: Mogrol can inhibit the synthesis of fatty acids by activating the expression level of AMPK signaling pathway related factors SREBP-1c and FASN, so as to play the role of regulating lipid metabolism.
[Abstract] Objective：To study the regulatory effect of mogrol (MO) on lipid metabolism of hepatic cancer cells and its molecular mechanism. Methods: Oleic acid (OA) was used to induce fat accumulation in hepatocellular carcinoma HepG2 cells and to establish a steatosis cell model. CCK-8 method was used to detect the cytotoxicity of MO to HepG2 cells, and an experimental working concentration without obvious cytotoxicity of MO was chosen. After being treated with different concentrations of MO, lipid accumulation in the cells was observed by oil red O staining, and the contents of triglyceride (TG) and cholesterol (TC) in the cells were measured. Key genes involving in lipid metabolism were screened out by high-throughput transcriptome sequencing qPCR was used to detect the mRNA expressions of ,SREBP-1c and FASN, while Western Blot was used to detect the protein expressions of p-AMPKα, SREBP-1c and FASN in cells of model group and treatment group. Results: After OA induction, a large amount of lipids accumulated in HepG2 cells, the contents of TG and TC increased significantly. Three key genes (SREBP-1c, FASN and p-AMPK α) involving in lipid metabolism of hepatic cancer cells were screened out. After OA induction, the mRNA expressions of SREBP-1c and FASN increased, the protein expression of p-AMPK α decreased while the protein expressions of SREBP-1c, FASN and other proteins increased significantly.After intervention with working concentration of MO, intracellular lipid accumulation, contents of TG and TC, mRNA expressions of SREBP-1c, FASN and protein expressions of SREBP-1c, FASN decreased significantly, while the expression of p-AMPKα increased.Conclusion: Mogrol can inhibit the synthesis of fatty acids by activating the expression level of AMPK signaling pathway related factors SREBP-1c and FASN, so as to play the role of regulating lipid metabolism.
2019, 26(8):882-887.
DOI: 10.3872/j.issn.1007-385X.2019.08.009
Abstract:
[Abstract] Objective: To investigate the expression of phosphorylated retinoblastoma (p-Rb) in advanced melanoma tissues and to analyze its correlation with clinical pathological characteristics and prognosis. Methods: The clinical data and paraffin-embedded tissue sections of 66 patients with advanced melanoma, who were diagnosed and treated in the Department of Renal Cancer and Melanoma of Peking University Cancer Hospital from 2011 to 2014, were collected. Expression of p-Rb in primary tumor tissues was detected by immunohistochemistry.Correlation analysis was performed on the relationship between the expression level of p-Rb in tumor tissues and clinical data such as clinicopathological features and overall survival. Results: The positive expression rate of p-Rb in advanced melanoma tissues was 57.6% (38/66). The positive rates of p-Rb in non-acral cutaneous type, acral type and mucosal type were 73.7% (14/19), 63.0% (17/27) and 35.0% (7/20), respectively, and the difference was statistically significant (P=0.039). The positive rate of p-Rb in different gene mutation subgroups was also different, with 83.3% (5/6) in the BRAF mutation group, 100.0% (2/2) in the c-KIT mutation group, and 100.0% in the N-RAS mutation group (9/9), 50.0% (1/2) in the PDGFRA mutation group, and 50.0% (1/2) in group with 2-gene mutation, and 44.4% (20/45) in the wild type gene group, and the difference was statistically significant (P=0.004). There was no correlation between p-Rb expression levels and age, gender, stage, ulcer, and serum LDH levels. The median overall survival (OS) of patients with positive p-Rb expression was slightly shorter than that of the patients with negative expression (30.0 vs 39.2 months), but the difference was not statistically significant (P=0.555). Conclusion: More than half of the advanced melanoma tissues have positive expression of p-Rb, and the positive rate of p-Rb in non-acral cutaneous type is higher than that of acral and mucosal types. And the positive rate of p-Rb in melanoma tissues of patients carrying c-KIT and N-RAS mutations is higher.
[Abstract] Objective: To investigate the expression of phosphorylated retinoblastoma (p-Rb) in advanced melanoma tissues and to analyze its correlation with clinical pathological characteristics and prognosis. Methods: The clinical data and paraffin-embedded tissue sections of 66 patients with advanced melanoma, who were diagnosed and treated in the Department of Renal Cancer and Melanoma of Peking University Cancer Hospital from 2011 to 2014, were collected. Expression of p-Rb in primary tumor tissues was detected by immunohistochemistry.Correlation analysis was performed on the relationship between the expression level of p-Rb in tumor tissues and clinical data such as clinicopathological features and overall survival. Results: The positive expression rate of p-Rb in advanced melanoma tissues was 57.6% (38/66). The positive rates of p-Rb in non-acral cutaneous type, acral type and mucosal type were 73.7% (14/19), 63.0% (17/27) and 35.0% (7/20), respectively, and the difference was statistically significant (P=0.039). The positive rate of p-Rb in different gene mutation subgroups was also different, with 83.3% (5/6) in the BRAF mutation group, 100.0% (2/2) in the c-KIT mutation group, and 100.0% in the N-RAS mutation group (9/9), 50.0% (1/2) in the PDGFRA mutation group, and 50.0% (1/2) in group with 2-gene mutation, and 44.4% (20/45) in the wild type gene group, and the difference was statistically significant (P=0.004). There was no correlation between p-Rb expression levels and age, gender, stage, ulcer, and serum LDH levels. The median overall survival (OS) of patients with positive p-Rb expression was slightly shorter than that of the patients with negative expression (30.0 vs 39.2 months), but the difference was not statistically significant (P=0.555). Conclusion: More than half of the advanced melanoma tissues have positive expression of p-Rb, and the positive rate of p-Rb in non-acral cutaneous type is higher than that of acral and mucosal types. And the positive rate of p-Rb in melanoma tissues of patients carrying c-KIT and N-RAS mutations is higher.
2019, 26(8):888-895.
DOI: 10.3872/j.issn.1007-385X.2019.08.010
Abstract:
[Abstract] Objective: To investigate the regulatory effect of lncRNA MALAT1/miR-141-3p/ZEB1 axis on the invasion, metastasis and epithelial mesenchymal transition (EMT) of gastric cancer (GC) cells. Methods: Thirty-eight pairs of GC tissues (non-necrotic part) and corresponding adjacent tissues (>5 cm away from tumor tissue) removed by general surgery in Wuhan Commercial Hospital from April 2014 to May 2017 were collected. Meanwhile, normal gastric epithelial GES1 cells and GC cell lines (SGC7901, HGC27,BGC823, MKN45 and MKN28) were selected. The expression level of MALAT1 and miR-141-3p in GC tissues and cell lines were detected by qPCR. The effect of MALAT1 knockdown on proliferation, migration and invasion of SGC7901 cells was determined by CCK-8 assay and Transwell assay. WB was performed for measuring the expression level of ZEB1, E-cadherin, N-cadherin and Vimentin.Dual luciferase reporter gene assay was used to validate the relationship among MALAT1, miR-141-3p and ZEB1. CCK-8 assay and Transwell assay were used to detect the effect of MALAT1/miR-141-3p/ZEB1 axis on biological behaviors of SGC7901 cells. Results:MALAT1 was over-expressed in GC tissues and cell lines (P<0.05 or P<0.01). Knockdown of MALAT1 significantly inhibited the proliferation, migration, invasion and EMT of SGC7901 cells (P<0.05 or P<0.01). The results of dual luciferase reporter gene assay showed that MALAT1 directly targeted miR-141-3p, as well as for miR-141-3p and ZEB1. Further experiment indicated that simultaneous over-expression of miR-141-3p and MALAT1 or ZEB1 could restore the biological behaviors of SGC7901 cells, which were inhibited by miR-141-3p. Conclusion: MALAT1 promotes the invasion, metastasis and EMT of GC SGC7901 cells by down-regulating the inhibitory effect of miR-141-3p on ZEB1.
[Abstract] Objective: To investigate the regulatory effect of lncRNA MALAT1/miR-141-3p/ZEB1 axis on the invasion, metastasis and epithelial mesenchymal transition (EMT) of gastric cancer (GC) cells. Methods: Thirty-eight pairs of GC tissues (non-necrotic part) and corresponding adjacent tissues (>5 cm away from tumor tissue) removed by general surgery in Wuhan Commercial Hospital from April 2014 to May 2017 were collected. Meanwhile, normal gastric epithelial GES1 cells and GC cell lines (SGC7901, HGC27,BGC823, MKN45 and MKN28) were selected. The expression level of MALAT1 and miR-141-3p in GC tissues and cell lines were detected by qPCR. The effect of MALAT1 knockdown on proliferation, migration and invasion of SGC7901 cells was determined by CCK-8 assay and Transwell assay. WB was performed for measuring the expression level of ZEB1, E-cadherin, N-cadherin and Vimentin.Dual luciferase reporter gene assay was used to validate the relationship among MALAT1, miR-141-3p and ZEB1. CCK-8 assay and Transwell assay were used to detect the effect of MALAT1/miR-141-3p/ZEB1 axis on biological behaviors of SGC7901 cells. Results:MALAT1 was over-expressed in GC tissues and cell lines (P<0.05 or P<0.01). Knockdown of MALAT1 significantly inhibited the proliferation, migration, invasion and EMT of SGC7901 cells (P<0.05 or P<0.01). The results of dual luciferase reporter gene assay showed that MALAT1 directly targeted miR-141-3p, as well as for miR-141-3p and ZEB1. Further experiment indicated that simultaneous over-expression of miR-141-3p and MALAT1 or ZEB1 could restore the biological behaviors of SGC7901 cells, which were inhibited by miR-141-3p. Conclusion: MALAT1 promotes the invasion, metastasis and EMT of GC SGC7901 cells by down-regulating the inhibitory effect of miR-141-3p on ZEB1.
2019, 26(8):896-903.
DOI: 10.3872/j.issn.1007-385X.2019.08.011
Abstract:
[Abstract] Objective: To investigate the molecular mechanisms of lncRNA XIST/miR-34a-5p/SIRT6 axis regulating the proliferation and metastasis of oral squamous cell carcinoma (OSCC) cells. Methods: Specimens of tumor tissues and paracancer tissues of 47 patients with OSCC, who visited the Qingdao Stomatological Hospital from March 2013 to March 2018, were enrolled in this study. qPCR was performed to measure the mRNA expressions of lncRNA XIST, miR-34a-5p and SIRT6 in OSCC tissues and cell lines. WB was performed to measure the protein levels of SIRT6, Ki67, pcDNA, cleaved-caspase3, cleaved-caspase8, E-cadherin and vimentin in OSCC tissues and cell lines. CCK-8 assay was performed to observe the effect of lncRNA XIST knockdown on proliferation of Cal-27 and Tca-8113 cells; Tanswell assay was performed to detect migration and invasion of Cal-27 and Tca-8113 cells; flow cytometry was performed to detect the apoptosis of Cal-27 and Tca-8113 cells; and dual luciferase reporter assay was performed to verify the relationship between lncRNA XIST and miR-34a, or miR-34a and SIRT6. Results: Expressions of lncRNA XIST and SIRT6 were up-regulated in OSCC tissues and cell lines (all P<0.05), reversely, miR-34a-5p was down-regulated in OSCC tissues and cell lines (P<0.01). lncRNA XIST knockdown significantly suppressed OSCC cells proliferation, migration and invasion, and induced apoptosis of OSCC cells (all P<0.01); however, simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect.lncRNA XIST knockdown significantly increased cell proliferation and metastasis related protein expression in OSCC cells (all P<0.01), and simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect. In addition, lncRNA XIST directly targeted miR-34a-5p, and miR-34a-5p targeted SIRT6, lncRNA XIST upregulated SIRT6 expression via inhibiting miR-34a-5p (P<0.01). Conclusion: Taken together, lncRNA XIST/miR-34a-5p/SIRT6 axis regulates the proliferation and metastasis of OSCC cells and provides potential therapeutic targets for OSCC.
[Abstract] Objective: To investigate the molecular mechanisms of lncRNA XIST/miR-34a-5p/SIRT6 axis regulating the proliferation and metastasis of oral squamous cell carcinoma (OSCC) cells. Methods: Specimens of tumor tissues and paracancer tissues of 47 patients with OSCC, who visited the Qingdao Stomatological Hospital from March 2013 to March 2018, were enrolled in this study. qPCR was performed to measure the mRNA expressions of lncRNA XIST, miR-34a-5p and SIRT6 in OSCC tissues and cell lines. WB was performed to measure the protein levels of SIRT6, Ki67, pcDNA, cleaved-caspase3, cleaved-caspase8, E-cadherin and vimentin in OSCC tissues and cell lines. CCK-8 assay was performed to observe the effect of lncRNA XIST knockdown on proliferation of Cal-27 and Tca-8113 cells; Tanswell assay was performed to detect migration and invasion of Cal-27 and Tca-8113 cells; flow cytometry was performed to detect the apoptosis of Cal-27 and Tca-8113 cells; and dual luciferase reporter assay was performed to verify the relationship between lncRNA XIST and miR-34a, or miR-34a and SIRT6. Results: Expressions of lncRNA XIST and SIRT6 were up-regulated in OSCC tissues and cell lines (all P<0.05), reversely, miR-34a-5p was down-regulated in OSCC tissues and cell lines (P<0.01). lncRNA XIST knockdown significantly suppressed OSCC cells proliferation, migration and invasion, and induced apoptosis of OSCC cells (all P<0.01); however, simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect.lncRNA XIST knockdown significantly increased cell proliferation and metastasis related protein expression in OSCC cells (all P<0.01), and simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect. In addition, lncRNA XIST directly targeted miR-34a-5p, and miR-34a-5p targeted SIRT6, lncRNA XIST upregulated SIRT6 expression via inhibiting miR-34a-5p (P<0.01). Conclusion: Taken together, lncRNA XIST/miR-34a-5p/SIRT6 axis regulates the proliferation and metastasis of OSCC cells and provides potential therapeutic targets for OSCC.
12 Progress and challenges of chimeric antigen receptor gene modified-T cell immunotherapy to cancer
2019, 26(8):904-909.
DOI: 10.3872/j.issn.1007-385X.2019.08.012
Abstract:
[摘要] 过继细胞疗法（ACT）的飞速发展使其成为肿瘤治疗手段中的一项新热点，其中嵌合抗原受体修饰的T细胞（CAR-T细胞）在治疗恶性血液肿瘤中取得的成果更是令人振奋，同时也为实体瘤的治疗提供了新策略。但是，目前CAR-T 细胞免疫疗法在肿瘤治疗过程中的局限性也日渐显露。本文旨在针对CAR-T细胞在肿瘤治疗中的研究进展及治疗中的挑战予以简要探讨。
[摘要] 过继细胞疗法（ACT）的飞速发展使其成为肿瘤治疗手段中的一项新热点，其中嵌合抗原受体修饰的T细胞（CAR-T细胞）在治疗恶性血液肿瘤中取得的成果更是令人振奋，同时也为实体瘤的治疗提供了新策略。但是，目前CAR-T 细胞免疫疗法在肿瘤治疗过程中的局限性也日渐显露。本文旨在针对CAR-T细胞在肿瘤治疗中的研究进展及治疗中的挑战予以简要探讨。
2019, 26(8):910-915.
DOI: 10.3872/j.issn.1007-385X.2019.08.013
Abstract:
[摘要] 现阶段传统的放化疗给胃癌带来了一定的生存获益，但胃癌总体生存率仍不理想。迫切需要继续挖掘新的治疗靶标和诊断治疗方案。密封蛋白（claudin）是细胞连接复合体中紧密连接的重要组成成分，其异常表达导致的紧密连接功能失常会促进肿瘤的增殖和转移。Claudin 家族蛋白如claudin1、4、7、18.2 等的突变和表达水平异常影响胃癌细胞的增殖、侵袭、转移和凋亡等，具有作为胃癌诊断、治疗和预后的生物标志物的巨大潜力。以靶向claudin18.2 的药物IMAB362 为代表，目前已开发出多种针对claudin 蛋白的靶向药物，相关的临床试验已在胃癌、胰腺癌、生殖细胞瘤以及卵巢癌等恶性肿瘤患者中开展，针对claudin 蛋白的研究可能有助于探寻胃癌的早期诊断和精准治疗的新策略。
[摘要] 现阶段传统的放化疗给胃癌带来了一定的生存获益，但胃癌总体生存率仍不理想。迫切需要继续挖掘新的治疗靶标和诊断治疗方案。密封蛋白（claudin）是细胞连接复合体中紧密连接的重要组成成分，其异常表达导致的紧密连接功能失常会促进肿瘤的增殖和转移。Claudin 家族蛋白如claudin1、4、7、18.2 等的突变和表达水平异常影响胃癌细胞的增殖、侵袭、转移和凋亡等，具有作为胃癌诊断、治疗和预后的生物标志物的巨大潜力。以靶向claudin18.2 的药物IMAB362 为代表，目前已开发出多种针对claudin 蛋白的靶向药物，相关的临床试验已在胃癌、胰腺癌、生殖细胞瘤以及卵巢癌等恶性肿瘤患者中开展，针对claudin 蛋白的研究可能有助于探寻胃癌的早期诊断和精准治疗的新策略。
14 Interaction between tumor cells and bone marrow stromal cells and its effect on tumor cell dormancy
2019, 26(8):916-920.
DOI: 10.3872/j.issn.1007-385X.2019.08.014
Abstract:
[摘要] 目前，癌症患者的存活率及无病生存率持续提高，但是肿瘤复发率依然较高。肿瘤复发与肿瘤细胞的休眠及耐药有着密不可分的内在联系。骨髓具有丰富的血管和营养，因此也是原发肿瘤远处转移的主要场所之一。当肿瘤细胞转移到骨髓内，可以与骨髓内的基质细胞相互作用，并触发肿瘤细胞休眠，休眠状态下的肿瘤细胞不仅可以存活很长的时间，而且对细胞毒性治疗顽强抵抗；最重要的是在特定的条件下，还可恢复增殖，最终引起肿瘤复发。其中，成骨细胞、间充质干细胞和部分细胞因子促进肿瘤细胞的休眠，破骨细胞和神经元细胞主要参与肿瘤细胞休眠的停止和转移的发生，而内皮细胞促进肿瘤细胞增殖或是休眠取决于周围环境的活化状态。本文就上述内容的研究现状做一简要综述。
[摘要] 目前，癌症患者的存活率及无病生存率持续提高，但是肿瘤复发率依然较高。肿瘤复发与肿瘤细胞的休眠及耐药有着密不可分的内在联系。骨髓具有丰富的血管和营养，因此也是原发肿瘤远处转移的主要场所之一。当肿瘤细胞转移到骨髓内，可以与骨髓内的基质细胞相互作用，并触发肿瘤细胞休眠，休眠状态下的肿瘤细胞不仅可以存活很长的时间，而且对细胞毒性治疗顽强抵抗；最重要的是在特定的条件下，还可恢复增殖，最终引起肿瘤复发。其中，成骨细胞、间充质干细胞和部分细胞因子促进肿瘤细胞的休眠，破骨细胞和神经元细胞主要参与肿瘤细胞休眠的停止和转移的发生，而内皮细胞促进肿瘤细胞增殖或是休眠取决于周围环境的活化状态。本文就上述内容的研究现状做一简要综述。
2019, 26(8):921-924.
DOI: 10.3872/j.issn.1007-385X.2019.08.015
Abstract:
[摘要] NKT细胞是一类特殊的T 淋巴细胞亚群，既表达NK 细胞受体，也表达T 细胞的相关受体。NKT细胞在多种免疫应答的调节中发挥重要作用，包括感染、自身免疫性疾病、代谢性疾病和癌症，其通过连接固有免疫系统和适应性免疫系统显示出强大的抗肿瘤活性。NKT细胞不仅能杀伤肿瘤细胞，也可激活其他抗肿瘤免疫细胞间接地发挥抗肿瘤作用，还能在肿瘤微环境中激活衰竭的免疫细胞，在抗肿瘤免疫中发挥重要的作用。本文就NKT细胞的生物学特性及其在抗肿瘤免疫中的作用作一综述。
[摘要] NKT细胞是一类特殊的T 淋巴细胞亚群，既表达NK 细胞受体，也表达T 细胞的相关受体。NKT细胞在多种免疫应答的调节中发挥重要作用，包括感染、自身免疫性疾病、代谢性疾病和癌症，其通过连接固有免疫系统和适应性免疫系统显示出强大的抗肿瘤活性。NKT细胞不仅能杀伤肿瘤细胞，也可激活其他抗肿瘤免疫细胞间接地发挥抗肿瘤作用，还能在肿瘤微环境中激活衰竭的免疫细胞，在抗肿瘤免疫中发挥重要的作用。本文就NKT细胞的生物学特性及其在抗肿瘤免疫中的作用作一综述。
2019, 26(8):925-927.
DOI: 10.3872/j.issn.1007-385X.2019.08.016
Abstract:
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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.