摘要
目的:探讨miR-101表达对上皮性卵巢癌耐药细胞SKOV3/DDP行为活性的影响及相关作用机制。方法:上调SKOV3/DDP细胞中miR-101表达水平。RT-PCR法检测SKOV3/DDP细胞中miR-101 mRNA表达水平,CCK-8检测SKOV3/DDP细胞的顺铂敏感性,划痕实验检测SKOV3/DDP细胞的迁移力,Transwell实验检测SKOV3/DDP细胞侵袭力,Annexin V-FITC/PI流式细胞实验检测SKOV3/DDP细胞凋亡率,Western blot法检测SKOV3/DDP细胞p-ERK1/2、波形蛋白(Vimentin)、钙粘蛋白E(E-cadherin)及Caspase-3蛋白表达水平。结果:与各对照组比较,miR-101组SKOV3/DDP细胞miR-101mRNA表达水平明显升高(P<0.05),顺铂敏感性明显增加,IC50显著降低(P<0.05);划痕距离明显增宽(P<0.05),透膜细胞数显著减少(P<0.05);细胞凋亡率显著增高(P<0.05),p-ERK1/2及Vimentin蛋白表达水平降低(P<0.05),E-cadherin及Caspase-3表达水平显著升高(P<0.05)。结论:高表达的miR-101可抑制SKOV3/DDP细胞中p-ERK1/2蛋白表达,同时抑制细胞行为活性,促进顺铂诱导的细胞凋亡。
Objective： To explore the effect of miR-101 on the behavioral activity of the chemo-resistant epithelial ovarian cancer cells SKOV3/DDP and the related mechanism.Methods： SKOV3/DDP cells were up-regulated the expression of miR-101. RT-PCR assay was applied for the expression of miR-101 mRNA,and the cisplatin sensitivity of SKOV3/DDP cells were measured by CCK-8.The migration ability of SKOV3/DDP cells were tested by scratch assay and the invasion ability of SKOV3/DDP cells were tested by Transwell assay.The apoptosis of SKOV3/DDP cells were detected by V-FITC/PI Annexin flow cytometry,the expressions of p-ERK1/2,Vimntin and E-cadherin protein were detected by Western blot. Results： Compared with the control groups,the expression level of miR-101 mRNA in miR-101 group was increased significantly（ P〈0. 05）,the cisplatin IC50 of cisplatin was significantly decreased versus the control cells（ P〈0. 05）,and the wound healing distance was significantly broadened（ P〈0.05）,and the number of membrane cells was significantly decreased（ P〈0.05）.The apoptosis rate of miR-101 plasmid group was much higher than that of control groups（ P〈0.05）.The expression levels of p-ERK1/2 and Vimntin protein in miR-101 group were significant lower than those in control groups（ P〈0.05）,while the E-cadherin and Caspase-3 expressions were obviously induced.Conclusion： Overexpression of miR-101 could inhibit the expression of p-ERK1/2 protein and inhibit the biological activity of SKOV3/DDP cells,and promote the apoptosis of SKOV3/DDP cell.
出处
《现代妇产科进展》
CSCD
北大核心
2017年第8期578-582,587,共6页
Progress in Obstetrics and Gynecology
基金
河北省临床医学优秀人才培养计划(No:310103020704)
