小干扰RNA介导的间皮素沉默促进顺铂诱导的卵巢癌耐药细胞SKOV3/cDDP的凋亡  

Small interference RNA-mediated mesothelin silencing promotes apoptosis of cisplatin-resistant SKOV3/cDDP cells

作　　者：刘丽萍[1] 吴小华 梁红霞[2] 

机构地区：[1]河北医科大学妇产科教研室,石家庄050000 [2]中国人民解放军白求恩国际和平医院妇产科,石家庄050082

出　　处：《肿瘤》2011年第9期777-782,共6页Tumor

基　　金：国家自然科学基金资助项目(编号:30572093);河北省自然科学基金资助项目(编号:C2005000804)

摘　　要：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 small interference RNA（siRNA）-mediated mesothelin（MSLN） silencing on the apoptosis of cisplatin-resistant SKOV3/cisplatin（cDDP）（ovarian carcinoma） cells.Methods：The expression level of MSLN protein in SKOV3/cDDP cells was determined by Western blotting.The RNA interference（RNAi） technique was used to interfere with the expression of MSLN in order to establish stably-infected SKOV3/cDDP-LV-MSLN-RNAi cells.The sensitivity of the SKOV3/cDDP-LV-MSLN-RNAi cells to cDDP was detected by MTT assay and colony formation assay.The apoptosis rate of SKOV3/cDDP-LV-MSLN-RNAi cells was examined by flow cytometry（FCM）.Results：The expression level of MSLN protein in the SKOV3/cDDP cells was higher than that in the SKOV3 cells（P=0.000）.The half inhibition concentration（IC50） of cDDP in the SKOV3/cDDP-LV-MSLN-RNAi cells（interference group） was lower than those in the SKOV3/cDDP-LV-MSLN-neg cells（without interference group） and the SKOV3/cDDP cells（blank control group）（P=0.000）.The inhibitory effect on colony formation was stronger and the apoptosis rate was higher in the interference group as compared with the other two groups（P〈0.05）.Conclusion：MSLN silencing enhances the apoptosis of cisplatin-resistant SKOV3/cDDP cells and reverses resistance to cDDP to some extent.MSLN probably plays an important role in resistance to cDDP in ovarian cancer.

关 键 词：卵巢肿瘤 基因沉默 RNA干扰 药物耐受性 细胞凋亡 间皮素 

分 类 号：R737.31[医药卫生—肿瘤]