论文部分内容阅读
目的:探讨小干扰RNA(small interference RNA,siRNA)介导的间皮素(mesothelin,MSLN)沉默对顺铂(cisplatin,cDDP)诱导的卵巢癌耐药细胞SKOV3/cDDP凋亡的影响。方法:应用蛋白质印迹法检测SKOV3/cDDP细胞MSLN蛋白的表达。通过RNA干扰(RNA interference,RNAi)技术干扰耐药细胞SKOV3/cDDP MSLN表达,建立稳定感染的细胞株SKOV3/cDDP-LV-MSLN-RNAi。应用MTT法和集落形成实验检测SKOV3/cDDP-LV-MSLN-RNAi细胞对cDDP的敏感度,FCM检测cDDP诱导的细胞凋亡。结果:SKOV3/cDDP细胞MSLN蛋白的表达水平明显高于SKOV3细胞(P=0.000);cDDP对干扰组SKOV3/cDDP-LV-MSLN-RNAi细胞的半数抑制浓度(half inhibition concentration,IC50)值明显低于阴性组SKOV3/cDDP-LV-MSLN-neg和空白组SKOV3/cDDP细胞(P=0.000);干扰组细胞的集落形成抑制率和凋亡率明显高于阴性组和空白组,差异有统计学意义(P<0.05)。结论:MSLN沉默可促进cDDP诱导的卵巢癌耐药细胞凋亡,在一定程度上逆转cDDP耐药。MSLN可能在卵巢癌cDDP耐药中发挥重要作用。
Objective: To investigate the effect of silencing mesothelin (MSLN) mediated by small interfering RNA (siRNA) on the apoptosis of SKOV3 / cDDP cells induced by cisplatin (cDDP). Methods: Western blotting was used to detect the expression of MSLN protein in SKOV3 / cDDP cells. The stably infected cell line SKOV3 / cDDP-LV-MSLN-RNAi was established by interfering the expression of MSLN in SKOV3 / cDDP cells by RNA interference (RNAi) technique. The sensitivity of SKOV3 / cDDP-LV-MSLN-RNAi cells to cDDP was detected by MTT and colony formation assay, and cDDP-induced apoptosis was detected by FCM. Results: The expression of MSLN protein in SKOV3 / cDDP cells was significantly higher than that in SKOV3 cells (P = 0.000). The IC50 value of cDDP in SKOV3 / cDDP-LV-MSLN-RNAi cells was significantly lower (P = 0.000). The inhibition rate and apoptosis rate of SKOV3 / cDDP-LV-MSLN-neg in the negative group and SKOV3 / cDDP cells in the blank group were significantly higher than those in the negative group and the blank group Significance (P <0.05). Conclusion: Silencing MSLN can promote cDDP-induced apoptosis of ovarian cancer cells and reverse cDDP resistance to a certain extent. MSLN may play an important role in ovarian cancer cDDP resistance.