LncRNA GATA3-AS1通过调控miR-362-3p/FABP5轴抑制宫颈癌细胞增殖、迁移及侵袭

LncRNA GATA3-AS1 inhibits the proliferation,migration and invasion of cervical cancer cells by regulating the miR-362-3p/FABP5 axis

目的:探究长链非编码RNA GATA3反义RNA 1(lncRNA GATA3-AS1)调控微小RNA-362-3p(miR-362-3p)表达对宫颈癌细胞恶性生物学行为的影响。方法:qRT-PCR检测宫颈癌细胞中lncRNA GATA3-AS1、miR-362-3p、FABP5表达;双荧光素酶报告基因实验验证lncRNA GATA3-AS1和miR-362-3p的靶向关系、miR-362-3p和FABP5的靶向关系;将细胞分为pcDNA-NC组、pcDNA-GATA3-AS1组、si-NC组、si-GATA3-AS1组、si-GATA3-AS1+inhibitor-NC组、si-GATA3-AS1+miR-362-3p inhibitor组、miR-NC组、miR-362-3p mimics组、miR-362-3p mimics+pcDNA-NC组、miR-362-3p mimics+pcDNA FABP5组;Western blot检测蛋白表达;EdU法检测细胞增殖;Transwell检测细胞迁移侵袭。结果:在宫颈癌细胞系中,GATA3-AS1、FABP5均为高表达,miR-362-3p均为低表达,选择HeLa细胞进行后续实验;双荧光素酶报告基因实验表明,lncRNA GATA3-AS1和miR-802、miR-362-3p和FABP5具有靶向关系;与pcDNA-NC组比较,pcDNA-GATA3-AS1组Hela细胞EdU阳性率、迁移侵袭及MMP-2、MMP-9表达明显上升(P<0.05);与si-NC组比较,si-GATA3-AS1组HeLa细胞EdU阳性率、迁移侵袭及MMP-2、MMP-9表达明显下降(P<0.05);抑制miR-362-3p表达或过表达FABP5均可以明显逆转沉默GATA3-AS1或过表达miR-362-3p对于HeLa细胞增殖、迁移、侵袭的抑制作用。结论:沉默GATA3-AS1可以靶向上调miR-362-3p表达,抑制FABP5表达,抑制宫颈癌HeLa细胞增殖迁移及侵袭。

Objective:To investigate the impacts of long non-coding RNA GATA3 antisense RNA 1(lncRNA GATA3-AS1)on the malignant biological behavior of cervical cancer cells by regulating the expression of microRNA-362-3p(miR-362-3p).Methods:qRT-PCR was applied to detect the expression of lncRNA GATA3-AS1,miR-362-3p,and FABP5 in cervical cancer cells.Double Luciferase reporter gene experiment was applied to verify the targeting relationship between lncRNA GATA3-AS1 and miR-362-3p,and the targeting relationship between miR-362-3p and FABP5.Cells were divided into pcDNA-NC group,pcDNA-GATA3-AS1 group,si-NC group,si-GATA3-AS1 group,si-GATA3-AS1+inhibitor NC group,si-GATA3-AS1+miR-362-3p inhibitor group,miR-NC group,miR-362-3p mimics group,miR-362-3p mimics+pcDNA-NC group,and miR-362-3p mimics+pcDNA FABP5 group.Western blot was applied to detect protein expression.EdU method was applied to detect cell proliferation.Transwell was applied to detect cell migration and invasion.Results:In cervical cancer cell lines,GATA3-AS1 and FABP5 were highly expressed,while miR-362-3p was low expressed,HeLa cells were selected for subsequent experiments.Double Luciferase reporter gene experiment showed that lncRNA GATA3-AS1 was targeted to miR-802,miR-362-3p was targeted to FABP5.Compared with the pcDNA-NC group,the EdU positive rate,migration and invasion,and the expression of MMP-2 and MMP-9 in Hela cells in the pcDNA-GATA3-AS1 group were obviously increased(P<0.05).Compared with the si-NC group,the EdU positive rate,migration and invasion,and the expression of MMP-2 and MMP-9 in HeLa cells in the si-GATA3-AS1 group obviously decreased(P<0.05).Inhibiting the expression of miR-362-3p or overexpressing FABP5 was able to obviously reverse the inhibitory effects of silencing GATA3-AS1 or overexpressing miR-362-3p on the proliferation,migration,and invasion of HeLa cells.Conclusion:Silencing GATA3-AS1 can targetingly up-regulate the expression of miR-362-3p,inhibit the expression of FABP5,and inhibit the proliferation,migration,and invasion of cervical cancer HeLa cells.