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Volume 25,Issue 12,2018 Table of Contents
2018, 25(12):1209-1217.
DOI: 10.3872/j.issn.1007-385X.2018.12.001
Abstract:
[Abstract] CAR-T cell therapy has developed rapidly in recent years, and has achieved amazing results in the treatment of some malignant tumors of the blood system, but little progress has been made in the treatment of solid tumors. At present, the main problems to be solved in CAR-T cell therapy are: (1) enhancing the killing activity of CAR-T cells; (2) relieving the immunosuppressive state of tumors;(3) bringing CAR-T cells into solid tumors; (4) enhancing the safety of CAR-T cell therapy. By optimizing the structure of CAR,a series of defects in the CAR-T cell therapy can be overcome, and the curative effect of CAR-T can be enhanced and the complications can be alleviated. In this paper, some optimization and improvement measures and methods on the structure design of CAR in recent years are elaborated, and the effectiveness and safety of the CAR-T cell therapy are explored.
[Abstract] CAR-T cell therapy has developed rapidly in recent years, and has achieved amazing results in the treatment of some malignant tumors of the blood system, but little progress has been made in the treatment of solid tumors. At present, the main problems to be solved in CAR-T cell therapy are: (1) enhancing the killing activity of CAR-T cells; (2) relieving the immunosuppressive state of tumors;(3) bringing CAR-T cells into solid tumors; (4) enhancing the safety of CAR-T cell therapy. By optimizing the structure of CAR,a series of defects in the CAR-T cell therapy can be overcome, and the curative effect of CAR-T can be enhanced and the complications can be alleviated. In this paper, some optimization and improvement measures and methods on the structure design of CAR in recent years are elaborated, and the effectiveness and safety of the CAR-T cell therapy are explored.
2018, 25(12):1218-1222.
DOI: 10.3872/j.issn.1007-385X.2018.12.002
Abstract:
[Abstract] Due to the spectacular therapy results in hematologic tumors, chimeric antigen receptor T (CAR-T) cell therapy has been the research hot-spot in the field of cell-immunotherapy. Viral vectors, as the critical raw material in CAR-T cell manufacturing, are closely related to the safety, efficacy and quality control of CAR-T cell products, in the aspects of the structure design of CAR gene, refinement of production process, quality control and setting of characterization and specification etc. Based on the research progress in lentiviral and γ-retroviral vectors development and the evaluation experiences, this paper discusses some common problems that need to be focused on in the preparation of viral vectors, expecting to provide references for the development and the authorization applications of domestic related products in the future.
[Abstract] Due to the spectacular therapy results in hematologic tumors, chimeric antigen receptor T (CAR-T) cell therapy has been the research hot-spot in the field of cell-immunotherapy. Viral vectors, as the critical raw material in CAR-T cell manufacturing, are closely related to the safety, efficacy and quality control of CAR-T cell products, in the aspects of the structure design of CAR gene, refinement of production process, quality control and setting of characterization and specification etc. Based on the research progress in lentiviral and γ-retroviral vectors development and the evaluation experiences, this paper discusses some common problems that need to be focused on in the preparation of viral vectors, expecting to provide references for the development and the authorization applications of domestic related products in the future.
2018, 25(12):1223-1229.
DOI: 10.3872/j.issn.1007-385X.2018.12.003
Abstract:
Objective: To prepare a new type of phycocyanin / carboxymethyl chitosan-CD55 ligand peptide (CPC /CMC-CD55sp)nanospheres, and to study its targeted therapeutic effect on cervical cancer Caski cells. Methods: The novel CPC/CMC-CD55sp nanospheres (CPC/CMC-CD55sp) were synthesized by ionic cross-linking method, and the properties of nanospheres were observed by transmission electron microscopy (DLS) and fourier transform infrared spectroscopy (FTIR). The expression of CD55 on the surface of Caski and fibroblast (L-929) cells was detected by Western blotting and flow cytometry. The effect of nanospheres on the proliferation of Caski cells was detected by CCK-8. Flow cytometry and fluorescence microscopy were used to detect the uptake of microspheres by Caski cells; Western blotting and flow cytometry were used to detect the effect of CPC/CMC-CD55sp on expressions of apoptosis-related proteins and apoptosis rate in Caski cells; the hemolysis test was used to determine the biological safety of the drug. Results: CPC/CMC-CD55sp was successfully prepared with good morphology and uniform diameter; and CD55 was highly expressed on the surface of Caski cells but low expressed on the surface of L-929 cells (P<0.01). CPC/CMC-CD55sp could targeted and efficiently reach Caski cells and be ingested into the cells. It exhibited weak hemolysis effect on human peripheral blood, which was in the safe range. CPC/CMC-CD55sp displayed obvious inhibitory effect on Caski cell proliferation, and could induce cell apoptosis (P<0.05 or P<0.01). Conclusion:The new CPC/CMC-CD55sp can targeted inhibit the growth of cervical cancer Caski cells via inducing its apoptosis and has good bio-safety, which provides a new idea for the research and development of anti-tumor marine drugs.
Objective: To prepare a new type of phycocyanin / carboxymethyl chitosan-CD55 ligand peptide (CPC /CMC-CD55sp)nanospheres, and to study its targeted therapeutic effect on cervical cancer Caski cells. Methods: The novel CPC/CMC-CD55sp nanospheres (CPC/CMC-CD55sp) were synthesized by ionic cross-linking method, and the properties of nanospheres were observed by transmission electron microscopy (DLS) and fourier transform infrared spectroscopy (FTIR). The expression of CD55 on the surface of Caski and fibroblast (L-929) cells was detected by Western blotting and flow cytometry. The effect of nanospheres on the proliferation of Caski cells was detected by CCK-8. Flow cytometry and fluorescence microscopy were used to detect the uptake of microspheres by Caski cells; Western blotting and flow cytometry were used to detect the effect of CPC/CMC-CD55sp on expressions of apoptosis-related proteins and apoptosis rate in Caski cells; the hemolysis test was used to determine the biological safety of the drug. Results: CPC/CMC-CD55sp was successfully prepared with good morphology and uniform diameter; and CD55 was highly expressed on the surface of Caski cells but low expressed on the surface of L-929 cells (P<0.01). CPC/CMC-CD55sp could targeted and efficiently reach Caski cells and be ingested into the cells. It exhibited weak hemolysis effect on human peripheral blood, which was in the safe range. CPC/CMC-CD55sp displayed obvious inhibitory effect on Caski cell proliferation, and could induce cell apoptosis (P<0.05 or P<0.01). Conclusion:The new CPC/CMC-CD55sp can targeted inhibit the growth of cervical cancer Caski cells via inducing its apoptosis and has good bio-safety, which provides a new idea for the research and development of anti-tumor marine drugs.
2018, 25(12):1230-1236.
DOI: 10.3872/j.issn.1007-385X.2018.12.004
Abstract:
Objective: To study the effects of kaempferol on invasion and migration of non-small cell lung cancer (NSCLC) A549 cells and the related mechanisms. Methods: CCK-8 was used to detect the effect of different concentrations of kaempferol on the proliferation of A549 cells. Transwell assay and wound healing assay were used to detect the ability of cell invasion and migration. The expressions of EMT-related proteins (N-cadherin, Snail-2 and E-cadherin) were detected by Western blotting. The effect of kaempferol on the mRNA and protein expressions of estrogen related receptor alpha (ERRα) was determined by qRT-PCR. ERRα over-expression vector (pLV-ERRα) was transfected into A549 cells, and the cell invasion, migration and the expression of EMT related proteins were detected by Transwell assay, wound healing assay and Western blotting, respectively. Results: Kaempferol dose-dependently inhibited the proliferation of A549 cells, and kaempferol at concentrations of 5, 10 and 20 μmol/L was used in the following experiments. After the treatment with different concentrations of kaempferol, the number of invasive cells and wound closure rate, the expression of N-cadherin and Snail-2, and the mRNA and protein levels of ERRα decreased significantly, while the expression of E-cadherin increased significantly,(all P<0.01). After transfection with pLV-ERRα , the number of invasive cells, scratch closure rate and the expressions of N-cadherin and Snail-2 of A549 cells over-expressing ERRα significantly increased, while the expression of E-cadherin decreased significantly (all P<0.05 or P<0.01); and all these indices were attenuated after the treatment with kaempferol (all P<0.05 or P<0.01). Conclusion:Kaempferol inhibits the invasion and migration as well as EMT of non-small cell lung cancer A549 cells by down-regulating ERRα, which may provide experimental basis for the clinical treatment of lung cancer.
Objective: To study the effects of kaempferol on invasion and migration of non-small cell lung cancer (NSCLC) A549 cells and the related mechanisms. Methods: CCK-8 was used to detect the effect of different concentrations of kaempferol on the proliferation of A549 cells. Transwell assay and wound healing assay were used to detect the ability of cell invasion and migration. The expressions of EMT-related proteins (N-cadherin, Snail-2 and E-cadherin) were detected by Western blotting. The effect of kaempferol on the mRNA and protein expressions of estrogen related receptor alpha (ERRα) was determined by qRT-PCR. ERRα over-expression vector (pLV-ERRα) was transfected into A549 cells, and the cell invasion, migration and the expression of EMT related proteins were detected by Transwell assay, wound healing assay and Western blotting, respectively. Results: Kaempferol dose-dependently inhibited the proliferation of A549 cells, and kaempferol at concentrations of 5, 10 and 20 μmol/L was used in the following experiments. After the treatment with different concentrations of kaempferol, the number of invasive cells and wound closure rate, the expression of N-cadherin and Snail-2, and the mRNA and protein levels of ERRα decreased significantly, while the expression of E-cadherin increased significantly,(all P<0.01). After transfection with pLV-ERRα , the number of invasive cells, scratch closure rate and the expressions of N-cadherin and Snail-2 of A549 cells over-expressing ERRα significantly increased, while the expression of E-cadherin decreased significantly (all P<0.05 or P<0.01); and all these indices were attenuated after the treatment with kaempferol (all P<0.05 or P<0.01). Conclusion:Kaempferol inhibits the invasion and migration as well as EMT of non-small cell lung cancer A549 cells by down-regulating ERRα, which may provide experimental basis for the clinical treatment of lung cancer.
2018, 25(12):1237-1243.
DOI: 10.3872/j.issn.1007-385X.2018.12.005
Abstract:
Objective: To investigate the relationship between miR-141-3p and transforming growth factorβ2 (TGF-β2), and its effects on the malignant biological behaviors of human prostate cancer cell line C4-2B. Methods: After the transfection of miR-141-3p mimic,the mRNA expression of miR-141-3p and TGF-β2 in C4-2B cells was detected by qRT-PCR. Bioinformatics method validated the relationship between miR-141-3p and TGF-β2. miR-141-3p mimic alone or with TGF-β2 over-expression vector was transfected into C4-2B cells, and then Western blotting was used to detect the expression of TGF- β2 protein in C4-2B cells, Hochest33258 staining was used to detect cell apoptosis, and Transwell assay was used to detect the invasion ability of cells in each group. Results: After the transfection of C4-2B cells with miR-141-3p mimic, the level of miR-141-3p increased significantly, and the level of TGF- β2 mRNA decreased significantly (all P<0.01). The activity of luciferase was significantly reduced after the co-transfection with miR-141-3p mimic and wild type report plasmid (P<0.01); However, the activity of luciferase was not obviously changed after co-transfection with miR-141-3p mimic and mutant type report plasmid (P>0.05). After co-transfection with miR-141-3p mimic and pc-TGF-β2, the proliferation of C4-2B cells decreased significantly, the number of apoptotic cells increased significantly, and the cell invasion ability decreased significantly (all P<0.01). Conclusion: miR-141-3p inhibits the proliferation and invasion of human prostate cancer C4-2B cells and induces cell apoptosis by targeting TGF-β2.
Objective: To investigate the relationship between miR-141-3p and transforming growth factorβ2 (TGF-β2), and its effects on the malignant biological behaviors of human prostate cancer cell line C4-2B. Methods: After the transfection of miR-141-3p mimic,the mRNA expression of miR-141-3p and TGF-β2 in C4-2B cells was detected by qRT-PCR. Bioinformatics method validated the relationship between miR-141-3p and TGF-β2. miR-141-3p mimic alone or with TGF-β2 over-expression vector was transfected into C4-2B cells, and then Western blotting was used to detect the expression of TGF- β2 protein in C4-2B cells, Hochest33258 staining was used to detect cell apoptosis, and Transwell assay was used to detect the invasion ability of cells in each group. Results: After the transfection of C4-2B cells with miR-141-3p mimic, the level of miR-141-3p increased significantly, and the level of TGF- β2 mRNA decreased significantly (all P<0.01). The activity of luciferase was significantly reduced after the co-transfection with miR-141-3p mimic and wild type report plasmid (P<0.01); However, the activity of luciferase was not obviously changed after co-transfection with miR-141-3p mimic and mutant type report plasmid (P>0.05). After co-transfection with miR-141-3p mimic and pc-TGF-β2, the proliferation of C4-2B cells decreased significantly, the number of apoptotic cells increased significantly, and the cell invasion ability decreased significantly (all P<0.01). Conclusion: miR-141-3p inhibits the proliferation and invasion of human prostate cancer C4-2B cells and induces cell apoptosis by targeting TGF-β2.
2018, 25(12):1244-1250.
DOI: 10.3872/j.issn.1007-385X.2018.12.006
Abstract:
Objective: To investigate the effect of tanshinone IIA on the invasion and migration of esophageal cancer EC9706 and KYSE70 cells, and to explore the underlying mechanism. Methods: Esophageal cancer cells (EC9706 and KYSE70) were divided into 4 groups: control group, tanshinone IIA groups (2, 4, 6 μg/ml). Cell prliferation viability was measured by CCK-8; Apoptosis was detected by flow cytometry; Invasion was tested by Transwell assay; And migration was measured by Scratch assay. The mRNA and protein levels of E-cadherin, Snail-2, Vimentin and N-cadherin were tested by quantitative Real-time reverse transcription PCR (qRT-PCR)and Western blotting, respectively. Results: Tanshinone IIA at concentrations less than 6 μg/ml did not affect the cell viability of esophageal cancer EC9706 and KYSE70 cells. The apoptosis in tanshinone IIA (4, 6 μg/ml) groups was significantly higher than that in control group (P< 0.01). The number of invasive cells per field and wound-healing rate in tanshinone IIA (2, 4, 6 μg/ml) groups were significantly lower than those in control group (all P<0.01). Moreover, the cell morphology was transformed from a spindle-shaped mesenchymal form into epithelial morphology after tanshinone IIA treatment. Compared with control group, the expression of E-cadherin in tanshinone IIA groups (2, 4, 6 μg/ml) was significantly up-regulated while the expressions of Snail-2, Vimentin and N-cadherin were significantly down-regulated (all P<0.01). Conclusion: Tanshinone IIA promotes apoptosis and attenuates the invasion and migration of esophageal cancer cells by inhibiting the epithelial-mesenchymal transition.
Objective: To investigate the effect of tanshinone IIA on the invasion and migration of esophageal cancer EC9706 and KYSE70 cells, and to explore the underlying mechanism. Methods: Esophageal cancer cells (EC9706 and KYSE70) were divided into 4 groups: control group, tanshinone IIA groups (2, 4, 6 μg/ml). Cell prliferation viability was measured by CCK-8; Apoptosis was detected by flow cytometry; Invasion was tested by Transwell assay; And migration was measured by Scratch assay. The mRNA and protein levels of E-cadherin, Snail-2, Vimentin and N-cadherin were tested by quantitative Real-time reverse transcription PCR (qRT-PCR)and Western blotting, respectively. Results: Tanshinone IIA at concentrations less than 6 μg/ml did not affect the cell viability of esophageal cancer EC9706 and KYSE70 cells. The apoptosis in tanshinone IIA (4, 6 μg/ml) groups was significantly higher than that in control group (P< 0.01). The number of invasive cells per field and wound-healing rate in tanshinone IIA (2, 4, 6 μg/ml) groups were significantly lower than those in control group (all P<0.01). Moreover, the cell morphology was transformed from a spindle-shaped mesenchymal form into epithelial morphology after tanshinone IIA treatment. Compared with control group, the expression of E-cadherin in tanshinone IIA groups (2, 4, 6 μg/ml) was significantly up-regulated while the expressions of Snail-2, Vimentin and N-cadherin were significantly down-regulated (all P<0.01). Conclusion: Tanshinone IIA promotes apoptosis and attenuates the invasion and migration of esophageal cancer cells by inhibiting the epithelial-mesenchymal transition.
2018, 25(12):1251-1258.
DOI: 10.3872/j.issn.1007-385X.2018.12.007
Abstract:
Objective: To explore the mechanism of miR-429 targeting PTEN to affect capecitabine-resistance in pancreatic cancer PANC-1 cells though the PI3K/AKT signaling pathway. Methods: Capecitabine-resistant pancreatic cancer cell line PANC-1/CAP was constructed, and the expression of miR-429 and PTEN were detected by quantitative Real-time polymerase chain reaction (qRT-PCR)and Western blotting. The effect of miR-429 knock-down on cell proliferation viability, apoptosis and capecitabine-resistance was measured by colony formation assay, CCK-8 assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. Subsequently,dual luciferase reporter assay verified that PTEN was a target gene of miR-429. Furthermore, the effect of miR-429 on PTEN-PI3K/AKT signaling pathway was measured by Western blotting. Results: miR-429 was found to be up-regulated in PANC-1 cells and PANC-1/CAP cells compared with the non-malignant pancreatic ductal cell line (HPDE6-C7) (P<0.05 or P<0.01). Moreover, silencing of miR-429 significantly decreased cell proliferation viability, capecitabine-resistance and enhanced apoptosis of PANC-1/CAP cells;additionally, dual luciferase reporter assay confirmed that PTEN was a target of miR-429 (P<0.05 or P<0.01). Suppression of miR-429 up-regulated PTEN and blocked the PI3K / AKT signaling pathway to decrease cell proliferation viability and further reduce the capecitabine-resistance of PANC-1/CAP cells (P<0.05 or P<0.01). Conclusion: miR-429/PTEN-PI3K/AKT signaling pathway plays a certain role in regulating the capecitabine-resistance of pancreatic cancer, and inhibition of miR-429 expression may reverse the resistance of PANC-1/CAP to capecitabine.
Objective: To explore the mechanism of miR-429 targeting PTEN to affect capecitabine-resistance in pancreatic cancer PANC-1 cells though the PI3K/AKT signaling pathway. Methods: Capecitabine-resistant pancreatic cancer cell line PANC-1/CAP was constructed, and the expression of miR-429 and PTEN were detected by quantitative Real-time polymerase chain reaction (qRT-PCR)and Western blotting. The effect of miR-429 knock-down on cell proliferation viability, apoptosis and capecitabine-resistance was measured by colony formation assay, CCK-8 assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. Subsequently,dual luciferase reporter assay verified that PTEN was a target gene of miR-429. Furthermore, the effect of miR-429 on PTEN-PI3K/AKT signaling pathway was measured by Western blotting. Results: miR-429 was found to be up-regulated in PANC-1 cells and PANC-1/CAP cells compared with the non-malignant pancreatic ductal cell line (HPDE6-C7) (P<0.05 or P<0.01). Moreover, silencing of miR-429 significantly decreased cell proliferation viability, capecitabine-resistance and enhanced apoptosis of PANC-1/CAP cells;additionally, dual luciferase reporter assay confirmed that PTEN was a target of miR-429 (P<0.05 or P<0.01). Suppression of miR-429 up-regulated PTEN and blocked the PI3K / AKT signaling pathway to decrease cell proliferation viability and further reduce the capecitabine-resistance of PANC-1/CAP cells (P<0.05 or P<0.01). Conclusion: miR-429/PTEN-PI3K/AKT signaling pathway plays a certain role in regulating the capecitabine-resistance of pancreatic cancer, and inhibition of miR-429 expression may reverse the resistance of PANC-1/CAP to capecitabine.
MA Yizhen
,
FAN Yuanyuan
,
NIE Xin
,
SUN Lili
,
ZHU Yilong
,
LI Yiquan
,
LI Wenjie
,
YIN Xunzhe
,
LI Shanzhi
,
ZHAO Jin
,
LI Xiao
,
GUO Yan
,
JIN Ningyi
2018, 25(12):1264-1269.
DOI: 10.3872/j.issn.1007-385X.2018.12.009
Abstract:
Objective: To investigate the inhibitory effect of recombinant oncolytic adenovirus Ad-Apoptin-hTERTp-E1A (ATV) on luciferase-labeled human lung cancer cells (A549-luc) and human lung cancer A549 cells, and to compare the differences in the inhibitory effect on two cell lines. Methods: ATV was used to infect A549-luc cells and A549 cells respectively. WST-1 and crystal violet staining were used to determine the difference in the inhibitory effect of ATV. Hoechst and Annexin V-FITC/PI staining were used to verify the inhibition mode of ATV. Results: WST-1 and crystal violet staining showed that ATV had significant inhibitory effect on both A549-luc and A549 cells (P<0.05). ATV showed significant inhibitory effect on both cells at 24, 48 and 72 h (P<0.05 or P<0.01), and reached the peak at 72 h; ATV at concentrations of 1, 10 and 100 MOI all showed inhibitory effect on both cells, and reached the peak at 100 MOI.Hoechst staining showed that A549-luc cells and A549 cells infected with ATV showed typical nuclear fragmentation and marginal set.The results of Annexin V-FITC/PI Flow cytometry showed that ATV infection resulted in apoptosis of A549-luc and A549 cells, which was in a time-dependent manner and reached the peak at 72 h(P<0.05 or P<0.01). Conclusion: Insertion of luciferase didn’t significant-ly change the inhibitory effect and inhibitory mode of ATV on A549-luc cells. ATV exerted its in vitro inhibitory effect on A549-luc and A549 cells by inducing cell apoptosis.
Objective: To investigate the inhibitory effect of recombinant oncolytic adenovirus Ad-Apoptin-hTERTp-E1A (ATV) on luciferase-labeled human lung cancer cells (A549-luc) and human lung cancer A549 cells, and to compare the differences in the inhibitory effect on two cell lines. Methods: ATV was used to infect A549-luc cells and A549 cells respectively. WST-1 and crystal violet staining were used to determine the difference in the inhibitory effect of ATV. Hoechst and Annexin V-FITC/PI staining were used to verify the inhibition mode of ATV. Results: WST-1 and crystal violet staining showed that ATV had significant inhibitory effect on both A549-luc and A549 cells (P<0.05). ATV showed significant inhibitory effect on both cells at 24, 48 and 72 h (P<0.05 or P<0.01), and reached the peak at 72 h; ATV at concentrations of 1, 10 and 100 MOI all showed inhibitory effect on both cells, and reached the peak at 100 MOI.Hoechst staining showed that A549-luc cells and A549 cells infected with ATV showed typical nuclear fragmentation and marginal set.The results of Annexin V-FITC/PI Flow cytometry showed that ATV infection resulted in apoptosis of A549-luc and A549 cells, which was in a time-dependent manner and reached the peak at 72 h(P<0.05 or P<0.01). Conclusion: Insertion of luciferase didn’t significant-ly change the inhibitory effect and inhibitory mode of ATV on A549-luc cells. ATV exerted its in vitro inhibitory effect on A549-luc and A549 cells by inducing cell apoptosis.
2018, 25(12):1270-1275.
DOI: 10.3872/j.issn.1007-385X.2018.12.010
Abstract:
Objective: To observe the expression of miR-488-5p in cervical cancer tissues and to explore its effect on the proliferation and migration of cervical cancer C33A cells. Methods: 12 pairs of cervical cancer tissues and corresponding paracancer tissues from patients,who underwent total hysterectomy at the Luoyang Central Hospital of Zhengzhou University from March 2017 to September 2017, were collected for this study; and the expression of miR-488-5p was detected by fluorescence quantitative and real-time polymerase chain reaction (qRT-PCR). Lipofectamine 3000 was used to transfect miR-488-5p (experiment group) and miR-NC (control group) into cervical cancer C33A cells. Cell cycle distribution was detected by Flow cytometry. Cell proliferation was assessed by CCK-8 assay and Transwell assay was used to detect cell migration. Bioinformatics software was used to predict the possible target genes of miR-488-5p, and luciferase activity assay was used to verify the binding of miR-488-5p to target genes. The expressions of tumor endothelial marker 8 (TEM8) and downstream EGFR signaling pathway related proteins in two groups were detected by qRT-PCR and Western blotting. Results: The relative expression level of miR-488-5p in cervical cancer tissues (1.33±0.20) was significantly lower than that in paracancer tissues (3.68±0.45) (P<0.01). The relative expression level of miR-488-5p in the experimental group (25.23±3.11)was significantly higher than that in the control group (1.02±0.10) (P<0.01). The percentage of C33A cells at G0/G1 phase in experimental group (53.39±2.48)% was significantly higher than that in control group (39.57±1.21)% (P<0.01). When the culture time extended to 96 h and 120 h, the proliferation ability of C33A cells in experimental group was significantly lower than that in control group (P<0.05), and the number of migrated cells in the experimental group (117.90±18.86) was significantly less than that in the control group (295.10±19.33) (P <0.01). Luciferase activity assay confirmed that miR-488-5p could directly bind with TEM8 and inhibit its expression (P<0.01). The relative expression of TEM8 mRNA in experimental group (0.42±0.06) was significantly lower than that in control group (1.00 ± 0.06) (P<0.01). After transfection with miR-488-5p for 48h, the protein expressions of TEM8, p-EGFR, p-ERK and p-AKT were significantly lower than those in control group (P <0.01). Conclusion: The expression of miR-488-5p in cervical cancer tissues was decreased. Over-expression of miR-488-5p could inhibit the cell cycle progression of cervical cancer cells and reduce the proliferation and migration of cervical cancer cells. The mechanism may be related to the interference of TEM8 gene expression.
Objective: To observe the expression of miR-488-5p in cervical cancer tissues and to explore its effect on the proliferation and migration of cervical cancer C33A cells. Methods: 12 pairs of cervical cancer tissues and corresponding paracancer tissues from patients,who underwent total hysterectomy at the Luoyang Central Hospital of Zhengzhou University from March 2017 to September 2017, were collected for this study; and the expression of miR-488-5p was detected by fluorescence quantitative and real-time polymerase chain reaction (qRT-PCR). Lipofectamine 3000 was used to transfect miR-488-5p (experiment group) and miR-NC (control group) into cervical cancer C33A cells. Cell cycle distribution was detected by Flow cytometry. Cell proliferation was assessed by CCK-8 assay and Transwell assay was used to detect cell migration. Bioinformatics software was used to predict the possible target genes of miR-488-5p, and luciferase activity assay was used to verify the binding of miR-488-5p to target genes. The expressions of tumor endothelial marker 8 (TEM8) and downstream EGFR signaling pathway related proteins in two groups were detected by qRT-PCR and Western blotting. Results: The relative expression level of miR-488-5p in cervical cancer tissues (1.33±0.20) was significantly lower than that in paracancer tissues (3.68±0.45) (P<0.01). The relative expression level of miR-488-5p in the experimental group (25.23±3.11)was significantly higher than that in the control group (1.02±0.10) (P<0.01). The percentage of C33A cells at G0/G1 phase in experimental group (53.39±2.48)% was significantly higher than that in control group (39.57±1.21)% (P<0.01). When the culture time extended to 96 h and 120 h, the proliferation ability of C33A cells in experimental group was significantly lower than that in control group (P<0.05), and the number of migrated cells in the experimental group (117.90±18.86) was significantly less than that in the control group (295.10±19.33) (P <0.01). Luciferase activity assay confirmed that miR-488-5p could directly bind with TEM8 and inhibit its expression (P<0.01). The relative expression of TEM8 mRNA in experimental group (0.42±0.06) was significantly lower than that in control group (1.00 ± 0.06) (P<0.01). After transfection with miR-488-5p for 48h, the protein expressions of TEM8, p-EGFR, p-ERK and p-AKT were significantly lower than those in control group (P <0.01). Conclusion: The expression of miR-488-5p in cervical cancer tissues was decreased. Over-expression of miR-488-5p could inhibit the cell cycle progression of cervical cancer cells and reduce the proliferation and migration of cervical cancer cells. The mechanism may be related to the interference of TEM8 gene expression.
10 Expression of HOPX in cervical cancer tissues and blood serum and its correlation to CEA and CA125
2018, 25(12):1276-1281.
DOI: 10.3872/j.issn.1007-385X.2018.12.011
Abstract:
Objective: To investigate the expression of HOPX gene in cervical cancer tissues and blood serum as well as its effect on cervical cancer HeLa cells, and to analyze its correlation to tumor maker CEA and CA125. Methods: 50 pairs of cervical cancer tissues and para-cancerous tissues as well as the peripheral blood samples from patients with cervical cancer, who were treated at Tianjin Binhai People’s Hospital and Tianjin Wuqing People’s Hospital from June 2015 to December 2017, were collected for this study; in addition,50 samples of blood serum from healthy people were used as control. Real-time quantitative PCR (qRT-PCR) and immumohistochemical staining (IHC) were used to detect mRNA and protein expressions of HOPX in tissue and serum samples, NCBI-GEO data base and TCGA data base were used to collect the information on HOPX gene and patients’prognosis, and the correlation between HOPX expression and patients’prognosis was analyzed. Vectors over-expressing HOPX or control vectors were transfected into HeLa cells; MTT assay and colony formation assay were used to examine the proliferation ability of HeLa cells, Tranwell assay was used to detect the migration and invasion of HeLa cells, and Western blotting was used to detect the expression of EMT-related proteins. Results:Both sample examination and data base information showed that the expression level of HOPX was down-regulated in tissue and serum samples of cervical cancer patients and was positively related with the survival of patients (r=0.736，P<0.05); while it’s expression was negatively related to the level of CEA and CA125 in cervical cancer tissues and serum (r=-0.678, P<0.05). HOPX over-expression inhibited cell proliferation, migration and invasion, promoted the expression of E-cadherin but inhibited the expression of Vimentin and ICAM1 (all P<0.05 or P<0.001). Conclusion: HOPX is low expressed in cervical cancer tissues and blood samples, and negatively correlated with CEA and CA125, but positively correlated with the survival of patients. Thus, combination of HOPX and CEA/CA125 may improve the early diagnosis rate of cervical cancer and provide a new strategy for precision treatment of cervical cancer in future.
Objective: To investigate the expression of HOPX gene in cervical cancer tissues and blood serum as well as its effect on cervical cancer HeLa cells, and to analyze its correlation to tumor maker CEA and CA125. Methods: 50 pairs of cervical cancer tissues and para-cancerous tissues as well as the peripheral blood samples from patients with cervical cancer, who were treated at Tianjin Binhai People’s Hospital and Tianjin Wuqing People’s Hospital from June 2015 to December 2017, were collected for this study; in addition,50 samples of blood serum from healthy people were used as control. Real-time quantitative PCR (qRT-PCR) and immumohistochemical staining (IHC) were used to detect mRNA and protein expressions of HOPX in tissue and serum samples, NCBI-GEO data base and TCGA data base were used to collect the information on HOPX gene and patients’prognosis, and the correlation between HOPX expression and patients’prognosis was analyzed. Vectors over-expressing HOPX or control vectors were transfected into HeLa cells; MTT assay and colony formation assay were used to examine the proliferation ability of HeLa cells, Tranwell assay was used to detect the migration and invasion of HeLa cells, and Western blotting was used to detect the expression of EMT-related proteins. Results:Both sample examination and data base information showed that the expression level of HOPX was down-regulated in tissue and serum samples of cervical cancer patients and was positively related with the survival of patients (r=0.736，P<0.05); while it’s expression was negatively related to the level of CEA and CA125 in cervical cancer tissues and serum (r=-0.678, P<0.05). HOPX over-expression inhibited cell proliferation, migration and invasion, promoted the expression of E-cadherin but inhibited the expression of Vimentin and ICAM1 (all P<0.05 or P<0.001). Conclusion: HOPX is low expressed in cervical cancer tissues and blood samples, and negatively correlated with CEA and CA125, but positively correlated with the survival of patients. Thus, combination of HOPX and CEA/CA125 may improve the early diagnosis rate of cervical cancer and provide a new strategy for precision treatment of cervical cancer in future.
2018, 25(12):1282-1289.
DOI: 10.3872/j.issn.1007-385X.2018.12.012
Abstract:
Objective: To investigate the expression of miR-1269 in non-small-cell lung cancer (NSCLC) tissues, and to explore its effect on the cellular biological characteristics of NSCLC A549 cells and the underlying mechanism. Methods: 34 pairs of NSCLC tissues and the corresponding adjacent para-cancerous tissues obtained from the patients, who underwent surgery in the Department of Breast Surgery, the Fourth Hospital of Hebei Medical University from Jan. 2017 to Jan. 2018, were collected for this study. The expression level of miR-1269 in above tissue specimens was examined by real-time fluorescent quantitative PCR. After transfection with miR-1269 mimics and mimics NC (negative control), the proliferation, migration and invasion of A549 cells were detected by MTS, Wound healing and Transwell assay, respectively; and the changes in cell cycle distribution of A549 cells were examined by flow cytometry.The bioinformatics tool was used to predict the possible target gene of miR-1269, and the regulation effect of miR-1269 on target gene was then validated by Western blotting and Dual-luciferase reporter assay. In the meanwhile, the protein expressions of cyclin depen-dent kinase inhibitor p21, Cyclin D2, and EMT-related proteins (E-cadherin and ZEB2) in the transfected A549 cells were measured by Western blotting. Results: The expression level of miR-1269 in NSCLC tissues was significantly higher than that in paracancerous tissues (2.81±2.27 vs 1.61±1.36, P<0.05). The capacities of proliferation, migration and invasion of A549 cells in miR-1269 mimics transfection group were significantly higher than those in mimics NC group and blank control group (all P<0.01). And the cell proportion at S-phase in miR-1269-mimics group was obviously higher than that in mimics NC group [(46.54±1.57)% vs (23.32±3.15)%，P<0.01].Bioinformatics analysis showed that miR-1269 could combine with 3’UTR of FOXO1 gene. After transfection with miR-1269 mimics,the expression level and luciferase activity of FOXO1 protein in A549 cells were significantly reduced (all P<0.01). Moreover, the protein expressions of p21 and E-cadherin were significantly decreased after over-expression of miR-1269 (all P<0.05), while the expressions of ZEB2 and Cyclin D2 were up-regulated (all P<0.05). Conclusion: The expression level of miR-1269 in NSCLC tissues was significantly increased, and it could enhance the proliferation, cell cycle progression, migration and invasion of A549 cells. The possible mechanism may be related to its targeted regulation of FOXO1.
Objective: To investigate the expression of miR-1269 in non-small-cell lung cancer (NSCLC) tissues, and to explore its effect on the cellular biological characteristics of NSCLC A549 cells and the underlying mechanism. Methods: 34 pairs of NSCLC tissues and the corresponding adjacent para-cancerous tissues obtained from the patients, who underwent surgery in the Department of Breast Surgery, the Fourth Hospital of Hebei Medical University from Jan. 2017 to Jan. 2018, were collected for this study. The expression level of miR-1269 in above tissue specimens was examined by real-time fluorescent quantitative PCR. After transfection with miR-1269 mimics and mimics NC (negative control), the proliferation, migration and invasion of A549 cells were detected by MTS, Wound healing and Transwell assay, respectively; and the changes in cell cycle distribution of A549 cells were examined by flow cytometry.The bioinformatics tool was used to predict the possible target gene of miR-1269, and the regulation effect of miR-1269 on target gene was then validated by Western blotting and Dual-luciferase reporter assay. In the meanwhile, the protein expressions of cyclin depen-dent kinase inhibitor p21, Cyclin D2, and EMT-related proteins (E-cadherin and ZEB2) in the transfected A549 cells were measured by Western blotting. Results: The expression level of miR-1269 in NSCLC tissues was significantly higher than that in paracancerous tissues (2.81±2.27 vs 1.61±1.36, P<0.05). The capacities of proliferation, migration and invasion of A549 cells in miR-1269 mimics transfection group were significantly higher than those in mimics NC group and blank control group (all P<0.01). And the cell proportion at S-phase in miR-1269-mimics group was obviously higher than that in mimics NC group [(46.54±1.57)% vs (23.32±3.15)%，P<0.01].Bioinformatics analysis showed that miR-1269 could combine with 3’UTR of FOXO1 gene. After transfection with miR-1269 mimics,the expression level and luciferase activity of FOXO1 protein in A549 cells were significantly reduced (all P<0.01). Moreover, the protein expressions of p21 and E-cadherin were significantly decreased after over-expression of miR-1269 (all P<0.05), while the expressions of ZEB2 and Cyclin D2 were up-regulated (all P<0.05). Conclusion: The expression level of miR-1269 in NSCLC tissues was significantly increased, and it could enhance the proliferation, cell cycle progression, migration and invasion of A549 cells. The possible mechanism may be related to its targeted regulation of FOXO1.
LIANG Jia
,
WU Xuan
,
KUANG Gang
,
REN Libing
,
SHEN Supeng
,
GUO Wei
,
GUO Yanli
,
ZHU Jingyun
,
DONG Zhiming
2018, 25(12):1290-1295.
DOI: 10.3872/j.issn.1007-385X.2018.12.013
Abstract:
Objective: To investigate the expression of long non-coding RNA NUP50-AS1 (lncRNA NUP50-AS1) in esophageal squamous cell carcinomas (ESCC) tissues and cell lines, and to explore its effect on proliferation, migration and invasion of human esophageal cancer Eca109 cells. Methods: 49 pairs of ESCC tissues and corresponding para-cancerous tissues obtained from the Biological Specimen Base of the Fourth Hospital of Hebei Medical University during Jan. 2015 to Jan. 2016 were used in this study. qRT-PCR method was applied to detect the expression of NUP50-AS1 in collected tissues samples and five esophageal cancer cell lines (TE1,TE13, Eca109, Kyse150 and Kyse170). ShRNAs were transiently transfected into Eca109 cells to interfere the expression of NUP50-AS1 gene, and finally, sh2-NUP50-AS1 was used for the following experiments. The effect of NUP50-AS1 gene knockdown on the proliferation of Eca109 cells was detected by MTS and colony formation assay; the effect of NUP50-AS1 gene knockdown on the migration of Eca109 cells was detected by scratch test, and the effect on cell invasion was detected by Transwell assay. Results: The expression of NUP50-AS1 in ESCC was correlated with the lymphnode metastasis and TNM stage (all P<0.01). The expression of NUP50-AS1 in ESCC tissues was significantly higher than that in corresponding normal tissues (2.003±0.870 vs 1.000±0.000, P<0.05). The expression of NUP50-AS1 in five esophageal cancer cell lines was significantly up-regulated (P<0.05), and it had the highest expression in Eca109 cell line. After transfection, sh2-NUP50-AS1 had the highest transfection efficiency, and knocking down NUP50-AS1 gene significantly inhibited the proliferation, invasion and migration of the Eca109 cells. Conclusion: The expression of lncRNA NUP50-AS1 in ESCC tissues was significantly higher than that in the para-cancerous tissues, and correlated with the TNM stage and lymphnode metastasis. The down-regulation of NUP50-AS1 inhibited the proliferation, invasion and migration of esophageal cancer cells.The high expression of NUP50-AS1 gene may be closely related to the occurrence and development of ESCC.
Objective: To investigate the expression of long non-coding RNA NUP50-AS1 (lncRNA NUP50-AS1) in esophageal squamous cell carcinomas (ESCC) tissues and cell lines, and to explore its effect on proliferation, migration and invasion of human esophageal cancer Eca109 cells. Methods: 49 pairs of ESCC tissues and corresponding para-cancerous tissues obtained from the Biological Specimen Base of the Fourth Hospital of Hebei Medical University during Jan. 2015 to Jan. 2016 were used in this study. qRT-PCR method was applied to detect the expression of NUP50-AS1 in collected tissues samples and five esophageal cancer cell lines (TE1,TE13, Eca109, Kyse150 and Kyse170). ShRNAs were transiently transfected into Eca109 cells to interfere the expression of NUP50-AS1 gene, and finally, sh2-NUP50-AS1 was used for the following experiments. The effect of NUP50-AS1 gene knockdown on the proliferation of Eca109 cells was detected by MTS and colony formation assay; the effect of NUP50-AS1 gene knockdown on the migration of Eca109 cells was detected by scratch test, and the effect on cell invasion was detected by Transwell assay. Results: The expression of NUP50-AS1 in ESCC was correlated with the lymphnode metastasis and TNM stage (all P<0.01). The expression of NUP50-AS1 in ESCC tissues was significantly higher than that in corresponding normal tissues (2.003±0.870 vs 1.000±0.000, P<0.05). The expression of NUP50-AS1 in five esophageal cancer cell lines was significantly up-regulated (P<0.05), and it had the highest expression in Eca109 cell line. After transfection, sh2-NUP50-AS1 had the highest transfection efficiency, and knocking down NUP50-AS1 gene significantly inhibited the proliferation, invasion and migration of the Eca109 cells. Conclusion: The expression of lncRNA NUP50-AS1 in ESCC tissues was significantly higher than that in the para-cancerous tissues, and correlated with the TNM stage and lymphnode metastasis. The down-regulation of NUP50-AS1 inhibited the proliferation, invasion and migration of esophageal cancer cells.The high expression of NUP50-AS1 gene may be closely related to the occurrence and development of ESCC.
13 Effect and mechanism of lncRNA HIT on cisplatin resistance in osteosarcoma tissues and U2OS cells
2018, 25(12):1296-1302.
DOI: 10.3872/j.issn.1007-385X.2018.12.014
Abstract:
Objective: To investigate the relationship between long non-coding RNA (lncRNA) HIT and cisplatin (DDP) resistance in osteosarcoma cells and the mechanism related to epithelial-mesenchymal transition (EMT). Methods: 42 pairs of osteosarcoma tissues and corresponding para-cancerous tissues (more than 5 cm away from the edge of cancer tissues) were collected at the Department of Orthopedics, Tianjin Hospital during June 2017 to June 2018. Quantitative Real-time PCR (qRT-PCR) was used to detect the mRNA expression of HIT and EMT related markers (Snail and E-cadherin) in the collected tissues. The DDP-resistant osteosarcoma U2OS cell line was constructed and human adrenal 293T cell line was used as control. Two sets of siRNA vectors targeting HIT loaded on lentivirus were transfected into cells with DDP-resistance as the interference group A and group B. Meanwhile, the U2OS cell line was transfected with HIT full-length vector and blank vector respectively, as over-expression group and blank group. The DDP 50% inhibitory concentration (IC50) was detected by MTT assay. qRT-PCR was used to detect the mRNA expressions of HIT, Snail and E-cadherin.Western blotting was used to detect the protein expressions of Snail and E-cadherin. RNA binding protein immunoprecipitation (RNAIP)assay was used to clarify the combination of HIT and Snail protein in the U2OS and 293T cells. Results: The mRNA expressions of HIT and Snail in osteosarcoma tissues were significantly higher than those in para-cancerous tissues, while the mRNA expression of Ecadherin was significantly lower than that in the paracancerous tissues. The mRNA expression of HIT and E-cadherin in osteosarcoma tissues was negatively correlated (all P<0.01). The DDP IC50 in the DDP-resistance group was significantly higher than that in the control group, interference group A and B, and the DDP IC50 in over-expression group was significantly higher than that in blank group (all P<0.01). The expression of HIT in resistance group was significantly higher than that in the control group, and the HIT expressions in interference group A and B were significantly lower than that in DDP-resistance and control group; moreover, the expression of HIT in over-expression group was significantly higher than that in blank group (all P<0.05 or P<0.01). The mRNA expression of Snail in DDPresistance group was significantly higher than that in the control group and interference group A and B, while the mRNA expression of E-cadherin in DDP-resistance group was significantly lower than that in the control group and interference group A and B; and the mRNA expression of E-cadherin in over-expression group was significantly lower than that in blank group. The protein expression of Snail in the DDP-resistance group was significantly higher than that in the control group and interference group A and B,while E-cadherin protein expression was significantly lower; and protein expression of Snail in over-expression group was significantly higher than that in blank group (all P<0.05 or P<0.01). The expression of HIT in the U20S and 293T cells treated by anti-Snail antibody induced by immunomagnetic beads was significantly higher than that in the cells treated by IgG antibody (P<0.01). Conclusion: HIT can promote EMT and cisplatin-resistance in osteosarcoma cells through up-regulation of Snail protein and inhibition of E-cadherin transcription activity.
Objective: To investigate the relationship between long non-coding RNA (lncRNA) HIT and cisplatin (DDP) resistance in osteosarcoma cells and the mechanism related to epithelial-mesenchymal transition (EMT). Methods: 42 pairs of osteosarcoma tissues and corresponding para-cancerous tissues (more than 5 cm away from the edge of cancer tissues) were collected at the Department of Orthopedics, Tianjin Hospital during June 2017 to June 2018. Quantitative Real-time PCR (qRT-PCR) was used to detect the mRNA expression of HIT and EMT related markers (Snail and E-cadherin) in the collected tissues. The DDP-resistant osteosarcoma U2OS cell line was constructed and human adrenal 293T cell line was used as control. Two sets of siRNA vectors targeting HIT loaded on lentivirus were transfected into cells with DDP-resistance as the interference group A and group B. Meanwhile, the U2OS cell line was transfected with HIT full-length vector and blank vector respectively, as over-expression group and blank group. The DDP 50% inhibitory concentration (IC50) was detected by MTT assay. qRT-PCR was used to detect the mRNA expressions of HIT, Snail and E-cadherin.Western blotting was used to detect the protein expressions of Snail and E-cadherin. RNA binding protein immunoprecipitation (RNAIP)assay was used to clarify the combination of HIT and Snail protein in the U2OS and 293T cells. Results: The mRNA expressions of HIT and Snail in osteosarcoma tissues were significantly higher than those in para-cancerous tissues, while the mRNA expression of Ecadherin was significantly lower than that in the paracancerous tissues. The mRNA expression of HIT and E-cadherin in osteosarcoma tissues was negatively correlated (all P<0.01). The DDP IC50 in the DDP-resistance group was significantly higher than that in the control group, interference group A and B, and the DDP IC50 in over-expression group was significantly higher than that in blank group (all P<0.01). The expression of HIT in resistance group was significantly higher than that in the control group, and the HIT expressions in interference group A and B were significantly lower than that in DDP-resistance and control group; moreover, the expression of HIT in over-expression group was significantly higher than that in blank group (all P<0.05 or P<0.01). The mRNA expression of Snail in DDPresistance group was significantly higher than that in the control group and interference group A and B, while the mRNA expression of E-cadherin in DDP-resistance group was significantly lower than that in the control group and interference group A and B; and the mRNA expression of E-cadherin in over-expression group was significantly lower than that in blank group. The protein expression of Snail in the DDP-resistance group was significantly higher than that in the control group and interference group A and B,while E-cadherin protein expression was significantly lower; and protein expression of Snail in over-expression group was significantly higher than that in blank group (all P<0.05 or P<0.01). The expression of HIT in the U20S and 293T cells treated by anti-Snail antibody induced by immunomagnetic beads was significantly higher than that in the cells treated by IgG antibody (P<0.01). Conclusion: HIT can promote EMT and cisplatin-resistance in osteosarcoma cells through up-regulation of Snail protein and inhibition of E-cadherin transcription activity.
2018, 25(12):1303-1307.
DOI: 10.3872/j.issn.1007-385X.2018.12.015
Abstract:
Objective: To observe the expression of long-chain non-coding RNA (lncRNA) RP3-340N1.2 in breast cancer tissues and its effect on proliferation and migration of breast cancer MCF-7 cells, and to explore the possible mechanism. Methods: 13 pairs of breast cancer tissues and adjacent tissues from breast cancer patients, who underwent radical surgery at the Cancer Center of the Affiliated People’s Hospital of Hubei University of Medicine from Jan. 2017 to Sep. 2017, were collected for this study. qRT-PCR was used to detect the differential expression of RP3-340N1.2 in collected tissue samples and breast cancer cell lines and normal breast epithelial cell line. RP3-340N1.2 plasmid (experimental group) and the negative control plasmid (control group) were transfected into breast cancer MCF-7 cells using Lipofectamine 3000. Cell counting (CCK-8) and Transwell migration assay were used to examine the effect of RP3-340N1.2 over-expression on proliferation and migration of MCF7 cells, the effect of RP3-340N1.2 over-expression on the mRNA expression of miR-134-5p and OPCML was detected by qRT-PCR, and Western blotting was used to detect the expression of OPCML protein. Results: The expression of RP3-340N1.2 in breast cancer tissues was significantly lower than that in adjacent tissues (P<0.01),and the expression of RP3-340N1.2 in breast cancer cell lines was significantly lower than that in normal breast epithelial cells (P<0.01). Up-regulation of RP3-340N1.2 decreased the proliferation and migration of MCF7 cells (all P<0.05). After over-expression of RP3-340N1.2 in MCF7 cells, the expression of miR-134-5p obviously decreased (P<0.01); moreover, the mRNA and protein expressions of OPCML significantly increased (P<0.01) while the expressions of cell cycle regulatory proteins (CDK4, Cyclin D2) and cell migration regulatory proteins (Vimentin and N-cadherin) decreased significantly (all P<0.01). Conclusion: RP3-340N1.2 is low expressed in breast cancer tissues and cell lines. Up-regulation of RP3-340N1.2 expression can lead to decreased expression of miR-134-5p and increased expression of OPCML gene, thereby inhibiting the proliferation and migration of breast cancer cells.
Objective: To observe the expression of long-chain non-coding RNA (lncRNA) RP3-340N1.2 in breast cancer tissues and its effect on proliferation and migration of breast cancer MCF-7 cells, and to explore the possible mechanism. Methods: 13 pairs of breast cancer tissues and adjacent tissues from breast cancer patients, who underwent radical surgery at the Cancer Center of the Affiliated People’s Hospital of Hubei University of Medicine from Jan. 2017 to Sep. 2017, were collected for this study. qRT-PCR was used to detect the differential expression of RP3-340N1.2 in collected tissue samples and breast cancer cell lines and normal breast epithelial cell line. RP3-340N1.2 plasmid (experimental group) and the negative control plasmid (control group) were transfected into breast cancer MCF-7 cells using Lipofectamine 3000. Cell counting (CCK-8) and Transwell migration assay were used to examine the effect of RP3-340N1.2 over-expression on proliferation and migration of MCF7 cells, the effect of RP3-340N1.2 over-expression on the mRNA expression of miR-134-5p and OPCML was detected by qRT-PCR, and Western blotting was used to detect the expression of OPCML protein. Results: The expression of RP3-340N1.2 in breast cancer tissues was significantly lower than that in adjacent tissues (P<0.01),and the expression of RP3-340N1.2 in breast cancer cell lines was significantly lower than that in normal breast epithelial cells (P<0.01). Up-regulation of RP3-340N1.2 decreased the proliferation and migration of MCF7 cells (all P<0.05). After over-expression of RP3-340N1.2 in MCF7 cells, the expression of miR-134-5p obviously decreased (P<0.01); moreover, the mRNA and protein expressions of OPCML significantly increased (P<0.01) while the expressions of cell cycle regulatory proteins (CDK4, Cyclin D2) and cell migration regulatory proteins (Vimentin and N-cadherin) decreased significantly (all P<0.01). Conclusion: RP3-340N1.2 is low expressed in breast cancer tissues and cell lines. Up-regulation of RP3-340N1.2 expression can lead to decreased expression of miR-134-5p and increased expression of OPCML gene, thereby inhibiting the proliferation and migration of breast cancer cells.
2018, 25(12):1308-1315.
DOI: 10.3872/j.issn.1007-385X.2018.12.016
Abstract:
[摘要] 肿瘤免疫治疗主要通过调节机体免疫和肿瘤之间的平衡来实现肿瘤治疗的目的，已证实对多种肿瘤具有显著的临床疗效，被认为是继手术、化疗、放疗后又一重要的治疗方法。但目前肿瘤免疫治疗尚无统一的临床应用方案，对不同的肿瘤或同一肿瘤的不同个体疗效差异巨大，严重制约其发展。既往研究发现,影响免疫检查点抑制剂反应和耐药性的关键因素包括肿瘤本身的特征（如癌症基因组、表观基因组和微环境）、肿瘤免疫表型、宿主免疫组分（全身免疫和抗肿瘤免疫）及其他的外部影响。然而，最新研究表明，肿瘤突变负荷、DNA修复基因、HLA基因型、PD-L1 表达以及肿瘤免疫抑制微环境与免疫检查点抑制剂的反应密切相关。因此，本文将从肿瘤突变负荷、DNA修复基因、HLA基因型、PD-L1 表达以及肿瘤免疫抑制微环境等5 个方面阐述其影响免疫检查点抑制剂的新机制，旨在为肿瘤的靶向治疗提供借鉴。
[摘要] 肿瘤免疫治疗主要通过调节机体免疫和肿瘤之间的平衡来实现肿瘤治疗的目的，已证实对多种肿瘤具有显著的临床疗效，被认为是继手术、化疗、放疗后又一重要的治疗方法。但目前肿瘤免疫治疗尚无统一的临床应用方案，对不同的肿瘤或同一肿瘤的不同个体疗效差异巨大，严重制约其发展。既往研究发现,影响免疫检查点抑制剂反应和耐药性的关键因素包括肿瘤本身的特征（如癌症基因组、表观基因组和微环境）、肿瘤免疫表型、宿主免疫组分（全身免疫和抗肿瘤免疫）及其他的外部影响。然而，最新研究表明，肿瘤突变负荷、DNA修复基因、HLA基因型、PD-L1 表达以及肿瘤免疫抑制微环境与免疫检查点抑制剂的反应密切相关。因此，本文将从肿瘤突变负荷、DNA修复基因、HLA基因型、PD-L1 表达以及肿瘤免疫抑制微环境等5 个方面阐述其影响免疫检查点抑制剂的新机制，旨在为肿瘤的靶向治疗提供借鉴。
2018, 25(12):1316-1321.
DOI: 10.3872/j.issn.1007-385X.2018.12.017
Abstract:
[摘要] 糖基化是生物体内蛋白质的基本修饰方式之一，通过影响蛋白质的折叠、运输和定位，从而参与人体多种生物学功能的调节。研究表明，异常糖基化修饰参与生物体内多种病理生理过程，包括恶性肿瘤和一些炎症性疾病，尤其与肿瘤的转移和侵袭密切相关。而上皮间质转化（epithelial-mesenchymal transition，EMT）指上皮细胞失去紧密连接转化为间质的复杂过程,是肿瘤转移的重要机制之一。本文主要对蛋白质糖。
[摘要] 糖基化是生物体内蛋白质的基本修饰方式之一，通过影响蛋白质的折叠、运输和定位，从而参与人体多种生物学功能的调节。研究表明，异常糖基化修饰参与生物体内多种病理生理过程，包括恶性肿瘤和一些炎症性疾病，尤其与肿瘤的转移和侵袭密切相关。而上皮间质转化（epithelial-mesenchymal transition，EMT）指上皮细胞失去紧密连接转化为间质的复杂过程,是肿瘤转移的重要机制之一。本文主要对蛋白质糖。
2018, 25(12):1322-1326.
DOI: 10.3872/j.issn.1007-385X.2018.12.018
Abstract:
[摘要] 蛋白精氨酸甲基化过程与蛋白的磷酸化、泛素化过程类似，是细胞中常见的翻译后修饰过程。精氨酸甲基转移酶(protein arginine methyltransferase, PRMT)是催化蛋白精氨酸残基氮原子发生甲基化的关键酶。PRMT5 能够催化产生对称性二甲基化精氨酸(symmetrically dimethylated arginine,sDMA)，参与调解生命活动。近年来，越来越多的研究表明，PRMT5 与肿瘤的发生发展及肿瘤转移密切相关，并将其作为新靶点进行抗肿瘤药物研究。本文旨在阐述PRMT5 与肿瘤发生发展的相关性和以PRMT5 为靶点的抗肿瘤药物开发及未来的研究发展方向。
[摘要] 蛋白精氨酸甲基化过程与蛋白的磷酸化、泛素化过程类似，是细胞中常见的翻译后修饰过程。精氨酸甲基转移酶(protein arginine methyltransferase, PRMT)是催化蛋白精氨酸残基氮原子发生甲基化的关键酶。PRMT5 能够催化产生对称性二甲基化精氨酸(symmetrically dimethylated arginine,sDMA)，参与调解生命活动。近年来，越来越多的研究表明，PRMT5 与肿瘤的发生发展及肿瘤转移密切相关，并将其作为新靶点进行抗肿瘤药物研究。本文旨在阐述PRMT5 与肿瘤发生发展的相关性和以PRMT5 为靶点的抗肿瘤药物开发及未来的研究发展方向。
2018, 25(12):1327-1332.
DOI: 10.3872/j.issn.1007-385X.2018.12.019
Abstract:
[摘要] 白细胞免疫球蛋白样受体亚家族B（leukocyte immunoglobulin-like receptor subfamily B，LILRB）在骨髓细胞、造血干细胞、神经细胞等多种机体细胞广泛表达。有研究发现，LILRB受体可以与多种配体结合并具有多种生物学功能，包括调节炎症反应、免疫耐受、细胞分化和神经系统的可塑性等。近年来研究发现，LILRB在多种实体瘤和血液系统肿瘤表达增高并与患者预后显著相关，同时LILRB与免疫抑制、肿瘤细胞生长和自我更新直接相关，具有肿瘤支持因子和免疫检查点分子的双重作用。此外，肿瘤细胞表达的LILRB与肿瘤微环境中的免疫细胞相互作用，调节机体对肿瘤的免疫反应，敲除小鼠的LILRB同源基因后，小鼠的正常造血功能未受到明显影响。上述研究结果提示，LILRBs 可能是肿瘤治疗的理想靶点。本文就LILRB与实体瘤和血液系统肿瘤的发生机制、LILRB在肿瘤细胞中的信号转导方式、LILRB与肿瘤的免疫治疗及需要解决的问题进行阐述，以期为后续的深入研究提供参考。
[摘要] 白细胞免疫球蛋白样受体亚家族B（leukocyte immunoglobulin-like receptor subfamily B，LILRB）在骨髓细胞、造血干细胞、神经细胞等多种机体细胞广泛表达。有研究发现，LILRB受体可以与多种配体结合并具有多种生物学功能，包括调节炎症反应、免疫耐受、细胞分化和神经系统的可塑性等。近年来研究发现，LILRB在多种实体瘤和血液系统肿瘤表达增高并与患者预后显著相关，同时LILRB与免疫抑制、肿瘤细胞生长和自我更新直接相关，具有肿瘤支持因子和免疫检查点分子的双重作用。此外，肿瘤细胞表达的LILRB与肿瘤微环境中的免疫细胞相互作用，调节机体对肿瘤的免疫反应，敲除小鼠的LILRB同源基因后，小鼠的正常造血功能未受到明显影响。上述研究结果提示，LILRBs 可能是肿瘤治疗的理想靶点。本文就LILRB与实体瘤和血液系统肿瘤的发生机制、LILRB在肿瘤细胞中的信号转导方式、LILRB与肿瘤的免疫治疗及需要解决的问题进行阐述，以期为后续的深入研究提供参考。
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- 《中国肿瘤生物治疗杂志》
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.