李 臻,郭春霞,王 佳,姚 彦,郭 莉,岳玉光.卵巢癌细胞中MTDH-PTEN互作对奥沙利铂耐药性的影响及机制[J].,2022,(14):2631-2635 |
卵巢癌细胞中MTDH-PTEN互作对奥沙利铂耐药性的影响及机制 |
The Effect and Mechanism of MTDH-PTEN Interaction on Oxaliplatin Resistance in Ovarian Cancer Cells |
投稿时间:2022-02-28 修订日期:2022-03-25 |
DOI:10.13241/j.cnki.pmb.2022.14.006 |
中文关键词: 卵巢癌细胞 MTDH-PTEN互作 奥沙利铂 耐药性 |
英文关键词: Ovarian cancer cells MTDH-PTEN interaction Oxaliplatin Drug resistance |
基金项目:陕西省科技计划项目(2020SF-030) |
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中文摘要: |
摘要 目的:探究卵巢癌细胞中MTDH-PTEN互作对奥沙利铂耐药性的影响及机制。方法:人卵巢癌细胞系通过慢病毒感染,将其分为对照组、NC-shRNA组和MTDH-shRNA组。通过RT-PCR分析不同组细胞MTDH和PTEN mRNA表达。通过MTT测定法测定细胞活力。通过CCK-8检测奥沙利铂诱导下的细胞增殖。通过膜联蛋白V染色细胞凋亡测定细胞凋亡。通过流式细胞仪分析细胞周期。通过Transwell试验检测细胞迁移和侵袭。通过流式细胞仪分析细胞周期。通过共免疫沉淀分析MTDH和PTEN的互联作用。结果:MTDH-shRNA组MTDH mRNA表达较NC-shRNA组和对照组降低(P<0.05),MTDH-shRNA组PTEN mRNA表达较NC-shRNA组和对照组升高(P<0.05)。当未添加奥沙利铂(浓度为0 μM),各组细胞活力比较无差异(P>0.05)。当奥沙利铂浓度为2 μM、4 μM和8 μM时,MTDH-shRNA组细胞活力较NC-shRNA组和对照组降低(P<0.05)。0 h,各组细胞增殖比较无差异(P>0.05),第24 h、48 h和72 h时,MTDH-shRNA组细胞增殖较NC-shRNA组和对照组降低(P<0.05)。MTDH-shRNA组细胞凋亡率较NC-shRNA组和对照组升高(P<0.05)。MTDH-shRNA组G1期细胞占比较NC-shRNA组和对照组升高(P<0.05),MTDH-shRNA组S/M期细胞占比较NC-shRNA组和对照组降低(P<0.05)。MTDH-shRNA组细胞迁移和侵袭数量较NC-shRNA组和对照组减少(P<0.05)。结论:MTDH在卵巢癌细胞中与PTEN共表达并与可与PTEN相互作用,因此可通过抑制MTDH表达、诱导PTEN表达可以恢复对卵巢癌细胞对奥沙利铂的药物敏感性。 |
英文摘要: |
ABSTRACT Objective: To explore the effect and mechanism of MTDH-PTEN interaction on oxaliplatin resistance in ovarian cancer cells. Methods: Human ovarian cancer cell lines were infected with lentivirus and divided into control group, NC-shRNA group and MTDH-shRNA group. The expression of MTDH and PTEN mRNA in different groups of cells was analyzed by RT-PCR. The cell viability was measured by the MTT assay. CCK-8 is used to detect cell proliferation induced by oxaliplatin. Apoptosis was determined by staining with annexin V. Analyze the cell cycle by flow cytometry. Detect cell migration and invasion by Transwell test. Analyze the cell cycle by flow cytometry. The interaction between MTDH and PTEN was analyzed by co-immunoprecipitation. Results: The expression of MTDH mRNA in the MTDH-shRNA group was lower than that of the NC-shRNA group and control group (P<0.05), and the expression of PTEN mRNA in the MTDH-shRNA group was higher than that of the NC-shRNA group and control group (P<0.05). When oxaliplatin was not added (at a concentration of 0 μM), there was no difference in cell viability in each group (P>0.05). When the concentration of oxaliplatin was 2 μM, 4 μM and 8 μM, the cell viability of the MTDH-shRNA group was lower than that of the NC-shRNA group and the control group (P<0.05). At 0h, there was no difference in cell proliferation in each group (P>0.05). At 24 h, 48 h and 72 h, cell proliferation in the MTDH-shRNA group was lower than that in the NC-shRNA group and the control group (P<0.05). The apoptosis rate of MTDH-shRNA group was higher than that of NC-shRNA group and control group (P<0.05). The proportion of cells in the G1 phase of the MTDH-shRNA group was higher than that of the NC-shRNA group and the control group (P<0.05), and the proportion of cells in the S/M phase of the MTDH-shRNA group was lower than that of the NC-shRNA group and the control group (P<0.05). The number of cell migration and invasion in the MTDH-shRNA group was lower than that in the NC-shRNA group and the control group (P<0.05). Conclusion: MTDH is co-expressed with PTEN and can interact with PTEN in ovarian cancer cells, so the drug sensitivity of ovarian cancer cells to oxaliplatin can be restored by inhibiting the expression of MTDH and inducing the expression of PTEN. |
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