MiR-1275促进宫颈癌Hela细胞恶性进展的机制研究

Effect of miR-1275 on the Growth of Cervical Cancer Hela Cells and its Mechanism

目的探讨miR-1275对宫颈癌Hela细胞生长影响及其机制。方法将体外培养的Hela细胞分为miR-NC组、miR-1275组、anti-miR-NC组和anti-miR-1275组,采用RT-PCR检测miR-1275的表达,CCK-8法评估Hela细胞增殖活力,Transwell小室法测定Hela细胞迁移和侵袭能力,流式细胞仪检测细胞凋亡率。生物信息学软件预测miR-1275靶基因,双荧光素酶报告基因实验验证miR-1275和FHIT的靶向关系,Western blot检测FHIT蛋白的表达。结果上调miR-1275表达显著增加Hela细胞活力、侵袭和迁移能力(P<0.05),降低细胞凋亡率(P<0.05),而下调miR-1275表达具有相反作用。miR-1275与FHIT 3’UTR存在互补的结合位点,FHIT是miR-1275的靶基因。上调miR-1275表达显著抑制FHIT蛋白表达(P<0.05),而下调miR-1275表达显著促进FHIT蛋白表达(P<0.05)。结论miR-1275可促进宫颈癌Hela细胞增殖、侵袭和迁移,并抑制细胞凋亡,其分子机制可能与靶向调控FHIT表达有关。

Objective To investigate the effect of miR-1275 on the growth of cervical cancer Hela cells and its mechanism.Methods Hela cells cultured in vitro were divided into miR-NC group,miR-1275 group,anti-miR-NC group and anti-miR-1275 Group.The expression of miR-1275 was detected by RT-PCR,Cck-8 assay was used to evaluate the proliferation activity of Hela cells was evaluated by CCK-8 assay,the migration and invasion ability of Hela cells was measured by Transwell assay,and cell apoptosis rate was detected by flow cytometry.Bioinformatics software predicted miR-1275 target genes,and dual luciferase reporter assays validated the targeting relationship between miR-1275 and FHIT,and the expression of FHIT protein was detected by Western blot.Results of miR-1275 up-regulation significantly increased Hela cell viability,invasion and migration abulity(P<0.05),and decreased apoptosis rate(P<0.05),while miR-1275 down-regulation had an opposite effect.BFHIT 3'UTR has a binding site that was complementary to miR-1275,and FHIT was the target gene of miR-1275.miR-1275 up-regulation significantly inhibited FHIT protein expression(P<0.05),while miR-1275 down-regulation significantly promoted FHIT protein expression(P<0.05).Conclusion miR-1275 can promote cervical cancer Hela cells proliferation,invasion and migration,and inhibit cell apoptosis.Its molecular mechanism may be related to the targeted regulation of FHIT expression.