miR-96-5p通过下调肌动蛋白结合蛋白1促进卵巢癌干细胞顺铂耐药性

miR-96-5p promotes cisplatin resistance in ovarian cancer stem cells by down-regulating profilin 1

目的 探索微小核糖核酸(miRNA)miR-96-5p通过调控肌动蛋白结合蛋白1(profilin 1,PFN1)对卵巢癌干细胞(SiHa细胞)顺铂耐药的分子机制。方法 采用RT-qPCR检测miR-96-5p表达水平;将SiHa细胞置于不同浓度顺铂的培养基中,构建顺铂耐药细胞SiHa/DDP;采用Western blot检测蛋白的表达水平;双荧光素酶报告实验验证miR-96-5p与PFN1的靶向关系;MTT、Transwell和流式细胞术检测细胞增殖、侵袭和凋亡;成球实验检测细胞的成球能力。结果 miR-96-5p在SiHa细胞中高表达,且在SiHa/DDP细胞中的表达最高;敲降miR-96-5p可抑制SiHa/DDP细胞增殖、侵袭和干细胞成球能力并促进细胞凋亡;PFN1是miR-96-5p的潜在靶基因,过表达miR-96-5p通过下调PFN1促进SiHa/DDP细胞增殖、侵袭和干细胞成球,抑制细胞凋亡。结论 miR-96-5p通过靶向调控PFN1,促进SiHa/DDP细胞增殖、侵袭及其干细胞成球并抑制凋亡,从而增强SiHa/DDP细胞对顺铂的耐药性。

Objective To explore the molecular mechanism of the effect of microRNA(miR-96-5p)on ovarian cancer stem cells cisplatin resistance through profilin 1(PFN1).Methods The expression level of miR-96-5p was detected by RT-qPCR.SiHa cells were cultured in medium supplemented with various concentrations of cisplatin to construct cisplatin resistant SiHa/DDP.The expression levels of protein were detected by Western blot.The targeting relationship between miR-96-5p and PFN1 was verified by the dual-luciferase reporter gene experiment.Proliferation, invasion and apoptosis levels of SiHa/DDP cells were measured by MTT,Transwell and flow cytometry.Cell blasting assay was used to detect the globing ability of cells.Results miR-96-5p was highly expressed in SiHa cells, and the highest expression level was found in SiHa/DDP cells.Knockdown of miR-96-5p could inhibit SiHa/DDP cells proliferation, invasion and promote cell apoptosis.PFN1 was a potential target gene of miR-96-5p.Overexpression of PFN1 could promote SiHa/DDP cell proliferation, invasion, stem cell pellet and inhibition of apoptosis.Conclusion miR-96-5p promotes proliferation, invasion, stem cell pellet of stem cells and inhibits apoptosis of SiHa/DDP cells by targeting negative regulation of PFN1,thereby enhancing the drug resistance of SiHa/DDP cells to cisplatin.