摘要
目的:探讨抑制氯离子通道-3(ClC-3)基因表达对顺铂(DDP)诱导的人卵巢癌SKOV3/DDP细胞凋亡的促进作用,为卵巢癌的治疗提供实验依据。方法:转染pSH1-siRNA-ClC-3重组质粒至人卵巢癌SKOV3/DDP细胞,Western blotting方法检测转染重组质粒对ClC-3蛋白表达的影响;MTT法检测细胞增殖率;Western blotting方法检测凋亡相关蛋白细胞色素C(Cyt-c)及Caspase-3活化片段蛋白表达。结果:与SKOV3细胞比较,SKOV3/DDP细胞中ClC-3蛋白表达水平增加(P<0.05);转染pSH1-siRNA-ClC-3重组质粒后,SKOV3/DDP细胞ClC蛋白表达水平明显降低;ClC-3-siRNA联合DDP作用后,SKOV3/DDP细胞增殖率下降(P<0.05),Cyt-c和Caspase-3活化片段蛋白表达水平增加(P<0.05)。结论:pSH1-siRNA-ClC-3可有效抑制SKOV3/DDP细胞ClC-3蛋白的表达,并促进DDP诱导SKOV3/DDP细胞凋亡。
Objective To observe the promoting effect of inhibiting chloride channel-3（ClC-3） gene expression on the apoptosis of human ovarian cancer SKOV3/DDP cells induced by cisplatin（DDP）,and to provide the experimental basis for treatment of ovarian cancer.Methods The recombinant plamid pSH1-siRNA-ClC-3 was transfected into SKOV3/DDP cells.The ClC-3 expression changes in SKOV3/DDP cells transfected with pSH1-SiRNA-ClC-3 was detected by Western blotting.The vitality of SKOV3/DDP cells was measured by MTT assay.The expression levels of apoptosis-related proteins cytochrome C（Cyt-C） and Cleaved Caspase-3 were determined by Western blotting.Results Compared with SKOV3 cells,the ClC-3 expression levels in SKOV3/DDP cells was increased（P0.05）.Transfection of pSH1-SiRNA-ClC-3 suppressed the expression of ClC-3 efficiently in SKOV3/DDP cells（P0.05）.The proliferation rate of SKOV3/DDP cells was decreased（P0.05）,and the cyt-C and Cleaved Caspase-3 expression were increased after treated with ClC-3-siRNA combined with DDP（P0.05）.Conclusion pSH1-SiRNA-ClC-3 could suppress the expression of ClC-3 efficiently and enhance the apoptosis induced by DDP in SKOV3/DDP cells.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2012年第6期1106-1109,共4页
Journal of Jilin University：Medicine Edition
基金
吉林省教育厅资助课题(吉教科合字2009第156号
吉教科合字2011第117号
吉教科合字2012第394号)
吉林省科技厅自然科学基金资助课题(201215103)
关键词
卵巢肿瘤
氯通道-3
顺铂
细胞凋亡
ovarian tumor
chloride channel-3
cisplatin
apoptosis
