摘要
目的探讨miR-296靶向成纤维细胞生长因子受体1(FGFR1)对卵巢癌细胞生长、迁移及侵袭的影响。方法体外培养人SK-OV-3细胞,并将细胞分为对照组、miR-296 NC组、miR-296 mimics组、miR-296 mimics+FGFR1 NC组、miR-296 mimics+FGFR1组。实时荧光定量PCR(qRT-PCR)检测细胞中miR-296、FGFR1 mRNA水平;细胞计数试剂盒8(CCK-8)检测细胞活力;Transwell法检测细胞迁移和侵袭能力;蛋白免疫印迹(WB)法检测增殖、迁移和侵袭蛋白的表达;双荧光素酶报告实验分析miR-296与FGFR1的靶向关系。结果与对照组比较,miR-296 mimics组miR-296表达显著升高,FGFR1 mRNA表达显著降低(P<0.05);与miR-296 mimics组比较,miR-296 mimics+FGFR1组miR-296表达差异无统计学意义(P>0.05),FGFR1 mRNA表达显著升高(P<0.05)。与对照组比较,miR-296 mimics组细胞活力、迁移和侵袭细胞数目、蛋白FGFR1、增殖蛋白β-catenin、CyclinD1、迁移蛋白MMP-2、侵袭蛋白N-cadherin表达显著降低,E-cadherin蛋白表达显著升高(P<0.05);与miR-296 mimics组比较,miR-296 mimics+FGFR1组细胞活力、迁移和侵袭细胞数目、蛋白FGFR1、β-catenin、CyclinD1、MMP-2、N-cadherin表达显著升高,E-cadherin蛋白表达显著降低(P<0.05);双荧光素酶报告实验表明miR-296和FGFR1存在靶向关系。结论miR-296可能通过靶向FGFR1抑制SK-OV-3细胞EMT途径,抑制细胞生长、迁移和侵袭。
Objective To investigate the effects of miR-296 on the growth,migration and invasion of ovarian cancer cells by targeting fibroblast growth factor receptor 1(FGFR1).Methods SK-OV-3 cells were cultured in vitro and divided into control group,miR-296 NC group,miR-296 mimics group,miR-296 mimics+FGFR1 NC group and miR-296 mimics+FGFR1 group.The levels of miR-296 and FGFR1 mRNA were detected by real-time fluorescence quantitative PCR(qRT-PCR).Cell viability was detected by cell counting kit 8(CCK-8).Cell migration and invasion were detected by Transwell assay.Western blotting(WB)was used to detect the expression of proliferation,migration and invasion protein.Double luciferase reporter assay was used to analyze the targeting relationship between miR-296 and FGFR1.Results Compared with those in the control group,the expression of miR-296 in miR-296 mimics group was significantly increased,while the expression of FGFR1 mRNA was significantly decreased(P<0.05).Compared with those in miR-296 mimics group,the expression of miR-296 in miR-296 mimics+FGFR1 group had no significant difference(P>0.05),but the expression of FGFR1 mRNA was significantly increased(P<0.05).Compared with those in the control group,the cell viability,the numbers of migration and invasion cells,the expression of FGFR1,β-catenin,CyclinD1,MMP-2 and N-cadherin in miR-296 mimics group were significantly decreased,while the expression of E-cadherin was significantly increased(P<0.05).Compared with those in miR-296 mimics group,the cell viability,the numbers of migration and invasion cells,the expression of FGFR1,β-catenin,CyclinD1,MMP-2 and N-cadherin in miR-296 mimics+FGFR1 group were significantly increased,while the expression of E-cadherin was significantly decreased(P<0.05).Dual luciferase reporter assay showed that there is a targeted relationship between miR-296 and FGFR1.Conclusion MiR-296 may inhibit the EMT pathway of SK-OV-3 cells by targeting FGFR1,and inhibit cell growth,migration and invasion.
作者
李忆东
李婷婷
赵珠峰
吴国忠
LI Yidong;LI Tingting;ZHAO Zhufeng;WU Guozhong(Department of Basic Medicine,School of International Medical Technology,Shanghai Shanda College,Shanghai 201209,China)
出处
《西部医学》
2023年第5期648-653,共6页
Medical Journal of West China
基金
国家自然科学基金项目(81172100)
上海杉达学院护理学重点学科重点立项课题(2019zz35-yj)。
