LncRNA NEAT1/miR-361-5p/TRPM7轴对宫颈癌细胞恶性生物学行为的影响

Effect of lncRNA NEAT1/miR-361-5p/TRPM7 axis on malignant biological behavior of cervical cancer cells

目的探讨长链非编码RNA(lncRNA)核富集转录本1(NEAT1)/miR-361-5p/瞬时受体电位通道7(TRPM7)轴对宫颈癌细胞增殖、迁移和侵袭的影响。方法使用RT-qPCR和Western blot检测正常宫颈上皮细胞End1/E6E7和宫颈癌细胞(Caski、HeLa和C33a)内lncRNA NEAT1、miR-361-5p和TRPM7的表达;将HeLa细胞分为si-NC组、si-lncRNA NEAT1组、miRNC组、miR-361-5p组、si-lncRNA NEAT1+anti-miR-NC组、si-lncRNA NEAT1+anti-miR-361-5p组、miR-361-5p+vector组、miR-361-5p+TRPM7组,采用RT-qPCR检测细胞中lncRNA NEAT1、miR-361-5p表达;使用MTT、Transwell检测细胞的增殖、迁移和侵袭;使用Western blot检测TRPM7、MMP-2、CyclinD1和MMP-9蛋白的表达;使用双萤光素酶报告实验验证lncRNA NEAT1与miR-361-5p、miR-361-5p和TRPM7的靶向关系。结果在宫颈癌细胞Caski、HeLa、C33a中lncRNA NEAT1、TRPM7蛋白表达升高,而miR-361-5p表达水平降低;敲低lncRNA NEAT1或过表达miR-361-5p可降低CyclinD1、MMP-2、MMP-9蛋白的表达及细胞活性,并减少细胞迁移、侵袭。lncRNA NEAT1靶向调控miR-361-5p,抑制miR-361-5p表达可减弱敲低lncRNA NEAT1对宫颈癌细胞增殖、迁移和侵袭的作用;miR-361-5p靶向调控TRPM7,上调TRPM7可减弱过表达miR-361-5p对宫颈癌细胞增殖、迁移和侵袭。结论lncRNA NEAT1可能通过调控miR-361-5p/TRPM7轴促进宫颈癌细胞增殖、迁移和侵袭。

Objective To investigate the effects of long non-coding RNA(lncRNA)NEAT1/miR-361-5p/transient receptor potential channel 7(TRPM7)molecular axis on malignant biological behavior.Methods The expressions of lncRNA NEAT1,miR-361-5p and TRPM7 in normal cervical epithelial cells End1/E6E7 and cervical cancer cells(Caski,HeLa,C33a)were detected by RTqPCR and Western blot.HeLa cells were divided into si-NC group,si-lncRNA NEAT1 group,miRNC group,miR-361-5p group,si-lncRNA NEAT1+anti-miR-NC group,si-lncRNA NEAT1+antimiR-361-5p group,miR-361-5p+vector group,and miR-361-5p+TRPM7 group.RT-qPCR was used to detect the expression of lncRNA NEAT1 and miR-361-5p;MTT and Transwell was used to detect cell proliferation,migration and invasion;the protein expressions of TRPM7,MMP-2,CyclinD1 and MMP-9 were detected by Western blot;and dual luciferase reporter assay were used to detect the targeting relationship between lncRNA NEAT1 and miR-361-5p,miR-361-5p and TRPM7.Results The relative expression level of lncRNA NEAT1 and TRPM7 protein in Caski,He-La and C33a cells were increased,while the relative expression level of miR-361-5p were decreased.Knocking down lncRNA NEAT1 or overexpression of miR-361-5p can decrease the expression and cell activity of CyclinD1,MMP-2 and MMP-9 proteins,and reduce the number of cell migration and invasion.lncRNA NEAT1 targeted miR-361-5p regulation,and inhibition of miR-361-5p can attenu-ate the effect of lncRNA NEAT1 knockdown on the proliferation,migration and invasion of cervical cancer cells.miR-361-5p targets TRPM7,and up-regulation of TRPM7 can attenuate the effect of overexpression of miR-361-5p on the proliferation,migration and invasion of cervical cancer cells.Conclusions IncRNA NEAT1 may promote cervical cancer cell proliferation,migration,and invasion by regulating the miR-361-5p/TRPM7 axis.