长春新碱通过lncRNA XIST/miR-485-5p调控宫颈癌细胞增殖、迁移及侵袭的分子机制  

The molecular mechanism of vincristine regulating the proliferation,migration and invasion of cervical cancer cells through lncRNA XIST/miR-485-5p

作　　者：刘宇[1] 李延 雷玉荣[1] Liu Yu;Li Yan;Lei Yurong(Department of Obstetrics and Gynecology,Yan'an People's Hospital,Shanxi 716000,China)

机构地区：[1]陕西省延安市人民医院妇产科,延安716000

出　　处：《广西医科大学学报》2021年第10期1949-1955,共7页Journal of Guangxi Medical University

摘　　要：目的:探讨长春新碱对宫颈癌细胞增殖、迁移、侵袭的影响及其对lncRNA XIST/miR-485-5p通路的调控作用。方法:采用不同浓度的长春新碱处理宫颈癌细胞HeLa;pcDNA-NC、pcDNA-lncRNA XIST、miR-NC、miR-485-5p mimics分别转染入HeLa细胞;pcDNA-NC、pcDNA-lncRNA XIST、miR-NC、miR-485-5p mimics、miR-NC+pcDNA-lncRNA XIST、miR-485-5p mimics+pcDNA-lncRNA XIST分别转染入HeLa细胞后加入长春新碱处理细胞。采用MTT实验检测细胞活力,Transwell小室实验检测细胞迁移及侵袭能力,qPCR法检测lncRNA XIST、miR-485-5p表达,双荧光素酶报告基因实验检测lncRNA XIST与miR-485-5p的靶向关系,Western blotting法检测CyclinD1、MMP2、MMP9蛋白表达。结果:与NC组比较,长春新碱不同剂量组细胞活力显著降低,迁移及侵袭细胞数减少,CyclinD1、MMP2、MMP9蛋白水平降低,lncRNA XIST表达水平降低,miR-485-5p表达水平升高(均P<0.05)。与pcDNA-NC组比较,pcDNA-lncRNA XIST组细胞活力及CyclinD1、MMP2、MMP9蛋白表达水平升高,迁移及侵袭细胞数增多(均P<0.05),而lncRNA XIST过表达可明显逆转长春新碱对HeLa细胞增殖、迁移及侵袭的作用。与miR-NC组比较,miR-485-5p组细胞活力及CyclinD1、MMP2、MMP9蛋白表达水平降低,迁移及侵袭细胞数减少(均P<0.05),而miR-485-5p过表达可增强长春新碱对HeLa细胞增殖、迁移及侵袭的作用。双荧光素酶报告基因实验证实lncRNA XIST可竞争性结合miR-485-5p。与miR-NC+pcDNA-lncRNA XIST+高剂量组比较,miR-485-5p+pcDNA-lncRNA XIST+高剂量组细胞活力及CyclinD1、MMP2、MMP9蛋白表达水平降低,迁移及侵袭细胞数减少(均P<0.05)。结论:长春新碱可通过调控lncRNA XIST/miR-485-5p而减弱宫颈癌细胞增殖、迁移及侵袭能力。Objective:To explore the effect of vincristine on the proliferation,migration and invasion of cervical cancer cells and its regulation of lncRNA XIST/miR-485-5p pathway.Methods:Different concentrations of vincristine were used to treat cervical cancer cells HeLa.pcDNA-NC,pcDNA-lncRNA XIST,miR-NC,and miR-485-5p mimics were transfected into HeLa cells.pcDNA-NC,pcDNA-lncRNA XIST,miR-NC,miR-485-5p mimics,miR-NC+pcDNA-lncRNA XIST,and miR-485-5p mimics+pcDNA-lncRNA XIST were transfected into HeLa cells and then treated with vincristine cell.MTT test was used to detect cell viability.Transwell chamber experiment was used to detect cell migration and invasion ability.qPCR was used to detect the expression of lncRNA XIST and miR-485-5p.The dual luciferase reporter gene experiment was used to detect the targeting relationship between lncRNA XIST and miR-485-5p.Western blotting was used to detect the expression levels of CyclinD1,MMP2,and MMP9 protein.Results:Compared with the NC group,the cell viability,the number of migrated and invasive cells,and the expression levels of CyclinD1,MMP2,MMP9 and lncRNA XIST of different doses of vincristine group were decreased,while the expression level of miR-485-5p was increased(P<0.05).Compared with the pcDNA-NC group,the cell viability,the number of migrated and invasive cells,and the protein levels of CyclinD1,MMP2,and MMP9 were increased in pcDNA-lncRNA XIST group(P<0.05).Overexpression of lncRNA XIST significantly reversed the effects of vincristine on the proliferation,migration and invasion of HeLa cells.Compared with the miR-NC group,the cell viability,the number of migrated and invasive cells,and the protein levels of CyclinD1,MMP2,and MMP9 were decreased in miR-485-5p group(P<0.05).miR-485-5p overexpression enhanced the effect of vincristine on the proliferation,migration and invasion of HeLa cells.lncRNA XIST directly targeted miR-485-5p.Compared with the miR-NC+pcDNA-lncRNA XIST+high-dose group,the cell viability,the number of migrated and invasive cells and the protein leve

关 键 词：长春新碱 lncRNA XIST miR-485-5p 宫颈癌 增殖 迁移 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]