摘要
目的:探讨信号分子β连环素(β-catenin)在多个卵巢癌耐药细胞系中的表达及其对顺铂耐药性的影响。方法:首先在2个卵巢癌耐药细胞系(C13K,SKOV-3),2个卵巢癌敏感细胞系(OV2008,A2780),A2780顺铂敏感株、A2780-cis顺铂耐药株,COC10顺铂敏感株、COC1-cis顺铂耐药株中检测β-catenin蛋白的水平。通过小干扰RNA(si RNA)介导的RNA干扰技术靶向沉默细胞中β-catenin的表达,实时荧光定量聚合酶链反应(real-time PCR)和蛋白质印迹法(Western blotting)验证β-catenin基因沉默效果。台盼蓝染色方法分析β-catenin基因沉默对细胞化疗药物顺铂敏感性的影响,流式细胞术检测顺铂处理后β-catenin基因沉默细胞的凋亡率。Western blotting筛选多个凋亡相关信号分子的改变。结果:β-catenin在2个卵巢癌耐药细胞系C13K,SKOV-3中蛋白水平较敏感细胞系明显上调。与亲本A2780和COC10细胞相比,在耐药株A2780-cis和COC1-cis中表达水平也明显增高。β-catenin si RNA能显著抑制细胞中β-catenin蛋白的水平。β-catenin基因沉默后,卵巢癌耐药细胞C13K,SKOV-3对顺铂敏感性显著增高;流式细胞术进一步分析发现β-catenin基因沉默可以促进顺铂介导的卵巢癌耐药细胞的细胞凋亡。Western blotting发现β-catenin基因沉默后,顺铂处理可以显著上调促凋亡分子p53和caspase-3蛋白水平。结论:β-catenin在卵巢癌耐药细胞系表达明显增高,β-catenin沉默可以促进卵巢癌耐药细胞对顺铂的敏感性,并同时促进了顺铂介导的细胞凋亡。
Objective：To investigate the expression of β-catenin in several human ovarian cancer cell lines and its effect on cisplatin resistance. Methods：The protein level of β-catenin in two cisplatin-resistant cell lines（C13K,SKOV-3）,two cisplatin-sensitive cell lines（OV2008,A2780）,A2780,A2780-cis,COC10 and COC1-cis was examined by real-time PCR and western blotting analysis. Expression of β-catenin in ovarian cancer cells was knockdown by si RNA transfection. The effect ofβ-catenin silencing on cisplatin resistance was examined by trypan blue staining and flowcytometry. The change of apoptosisrelated proteins were examined by western blotting analysis. Results：The expression of β-catenin was upregulated in cisplatinresistant cell lines（C13K,SKOV-3）compared with cisplatin-sensitive cell lines. The expression of β-catenin in cisplatin-resistant A2780 and COC1 was also higher than that in paretnal A2780 and COC1 cells. si RNA targeting β-catenin effectively knockdown expression of β-catenin in ovarian cancer cells. Silencing of β-catenin enhanced the sensitivity of cisplatin-resistant cells（C13K,SKOV-3）to cisplatin. β-catenin si RNA also resulted in more apoptosis compared with control si RNA. Cisplatin treatment in β-catenin silencing cells induced the expression of apoptosis-related protein p53 and caspase-3. Conclusions：These finding suggested that β-catenin was upregulated in cisplatin-resistant cells. Gene silencing of β-catenin enhanced sensitivity to cisplatin in cisplatin-resistant cells by inducing more cisplatin-mediated apoptosis.
出处
《国际妇产科学杂志》
CAS
2015年第1期108-111,121,共5页
Journal of International Obstetrics and Gynecology
关键词
卵巢肿瘤
肿瘤细胞系
Β连环素
顺铂
抗药性
肿瘤
Ovarian neoplasms
Cell line
tumor
Beta catenin
Cisplatin
Drug resistance
neoplasm
