LncRNA CASC2与miR-155-5p的靶向关系及对子宫内膜癌细胞迁移、侵袭能力的影响

Effects of LncRNA CASC2 targeting miR-155-5p on migration and invasion of endometrial cancer cells

目的:探讨LncRNA癌易感性候选基因2(CASC2)对子宫内膜癌细胞迁移、侵袭能力的影响及其机制。方法:qRT-PCR法检测人正常子宫内膜上皮细胞hEEC,人子宫内膜癌细胞HEC-1A、KLE、HHUA中CASC2、miR-155-5p的表达。利用双荧光素酶报告实验验证CASC2和miR-155-5p的靶向关系。将HEC-1A细胞分别转染pcDNA3.1、pcDNA3.1-CASC2,anti-miR-NC、anti-miR-155-5p,pcDNA3.1、pcDNA3.1-CASC2+miR-NC、pcDNA3.1-CASC2+miR-155-5p。48 h后Western blot法测细胞中MMP-2、MMP-9蛋白的表达,Transwell小室实验检测细胞迁移和侵袭能力。结果:与hEEC细胞相比,HEC-1A、KLE、HHUA中CASC2表达降低,miR-155-5p表达升高(P<0.05)。双荧光素酶报告实验结果显示,CASC2和miR-155-5p存在靶向关系。与未转染组和pcDNA3.1组相比,pcDNA3.1-CASC2组迁移细胞数和侵袭细胞数均降低,MMP-2、MMP-9蛋白表达亦降低(P<0.05)。与未转染组和anti-miR-NC组相比,anti-miR-155-5p组细胞中miR-155-5p表达降低,迁移细胞数和侵袭细胞数均降低,MMP-2、MMP-9蛋白表达亦降低(P<0.05)。与pcDNA3.1-CASC2和pcDNA3.1-CASC2+miR-NC组相比,pcDNA3.1-CASC2+miR-155-5p组细胞中CASC2表达降低,miR-155-5p表达升高,迁移细胞数和侵袭细胞数均升高,MMP-2、MMP-9蛋白表达水平亦升高(P<0.05)。结论:CASC2可靶向负调节miR-155-5p,抑制子宫内膜癌细胞的迁移和侵袭。

Aim:To investigate the effects of LncRNA cancer susceptibility candidate 2(CASC2)on migration and invasion of endometrial cancer cells and its mechanism.Methods:The expressions of CASC2 and miR-155-5p in human normal endometrial epithelial cell hEEC,human endometrial cancer HEC-1A,KLE and HHUA cells were detected by qRT-PCR.The targeting relationship between CASC2 and miR-155-5p was verified by double luciferase report assay.HEC-1A cells were transfected with pcDNA3.1,pcDNA3.1-CASC2,anti-miR-NC,anti-miR-155-5p,pcDNA3.1,pcDNA3.1-CASC2+miR-NC,pcDNA3.1-CASC2+miR-155-5p,respectively for 48 hours.The expressions of MMP-2 and MMP-9 proteins were detected by Western blot,Transwell assay was used to detect cell migration and invasion ability.Results:Compared with hEEC,the CASC2 expression in HEC-1A,KLE and HHUA cells significantly decreased,and the miR-155-5p expression significantly increased(P<0.05).Double luciferase report assay showed that there was negative targeting relationship between CASC2 and miR-155-5p.Compared with the cells without transfection or transfected with pcDNA3.1,the number of migration cells invasion cells,and the expressions of MMP-2 and MMP-9 proteins in pcDNA3.1-CASC2 group were lower(P<0.05).Compared with the cells without transfection or transfected with anti-miR-NC,the number of migration cells,invasion cells,and the expressions of MMP-2 and MMP-9 proteins in anti-miR-155-5p group were lower(P<0.05).Compared with the cells transfected with pcDNA3.1-CASC2 or pcDNA3.1-CASC2+miR-NC,the expression of CASC2 was lower,that of miR-155-5p was higher in pcDNA3.1-CASC2+miR-155-5p group(P<0.05),the number of migration cells,invasion cells,and the expressions of MMP-2 and MMP-9 proteins were higher(P<0.05).Conclusions:CASC2 may inhibit the migration and invasion ability of endometrial cancer cells by negatively regulating miR-155-5p.