沉默间皮素基因对逆转卵巢癌SKOV3/cDDP细胞株顺铂耐药的影响

Reversal of Cisplatin Resistance in Ovarian Cancer SKOV3/cDDP Cell Line by Mesothelin Silence

【目的】探讨沉默间皮素(MSLN)基因对逆转卵巢癌顺铂(cDDP)耐药细胞株SK0V3/cDDP的影响。【方法】分别采用荧光定量PCR和Western blotting方法检测SKOV3/cDDP细胞及SKOV3细胞中MSLN mRNA和MSLN蛋白表达水平。分别用MSLN RNAi慢病毒液、空载体慢病毒液转染SKOV3/cD-DP细胞,建立稳定感染的细胞株SKOV3/cDDP-LV-MSLN-RNAi(实验组)和SKOV3/DDP-MSLN-neg(阴性组)。分别应用MTT法和FCM检测转染后耐药细胞对cDDP的敏感性及cDDP诱导的细胞凋亡情况。【结果】SKOV3/cDDP细胞MSLN mRNA和MSLN蛋白的表达水平明显高于敏感SKOV3细胞(P<0.01);转染间皮素RNAi慢病毒液的细胞MSLN蛋白的表达量明显低于空栽体慢病毒液转染的细胞和未进行任何转染操作的细胞,且差异有显著性(P<0.01)。实验组细胞的半数抑制浓度(IC50)值明显低于阴性组和空白组(P<0.01);实验组SKOV3/cDDP-LV-MSLN-RNAi细胞凋亡率明显高于阴性组和空白组(P<0.01)。【结论】MSLN沉默可增强卵巢癌SKOV3/cDDP细胞对cDDP的敏感性,促进cDDP诱导的卵巢癌耐药细胞凋亡,在一定程度上可逆转卵巢癌SKOV3/cDDP细胞株顺铂耐药。

【Objective】To investigate the effects and mechanisms of mesothelin silencing(MSLN)on reversing the cisplatin resistance of ovarian cancer drug cisplatin-resistant SKOV3/DDP cell line.【Methods】The expression level of MSLN mRNA and MSLN protein in SKOV3/cDDP cells and SKOV3 cells were detected by fluorescence quantitative PCR and Western blotting,respectively.SKOV3/cDDP cells were transfected with mesothelin RNAi lentivirus solution and empty vector lentivirus solution,respectively,to establish a stable cell line SKOV3/cDDP-LV-MSLN-RNAi(the experimental group),SKOV3/DDP-MSLN-neg(the negative control group).The sensitivity of the SKOV3/cDDP-LV-MSLN-RNAi cells to cDDP was detected by MTT assay.The apoptosis rate of SKOV3/cDDP-LV-MSLN-RNAi cells was examined by flow cytometry(FCM).【Results】The expression level of MSLN mRNA and MSLN protein in SKOV3/cDDP cells were significantly higher than that in sensitive SKOV3 cells(P<0.01).The expression of MSLN protein in cells with transfected mesothelin RNAi lentivirus solution was significantly lower than that in cells with transfected empty vector lentivirus solution and cells without any transfection.The difference was statistically significant(P<0.01).The half inhibition concentration(IC50)of the cells in the experimental group was significantly lower than that in the negative control group and the blank group(P<0.01).The apoptotic rate of SKOV3/cDDP-LV-MSLN-RNAi cells in the experimental group was significantly higher than that in the negative control group and the blank group,and the difference was statistically significant.【Conclusion】MSLN silence can enhance the sensitivity of ovarian cancer SKOV3/cDDP cells to cDDP,promote the apoptosis of drug-resistant ovarian cancer cells induced by cDDP,and to a certain degree reverse the cisplatin resistance of ovarian cancer SKOV3/cDDP cell lines.