lncRNA CBR3-AS1靶向调控miR-5195-3p促进胃癌细胞增殖、迁移及侵袭

lncRNA CBR3-AS1 promotes proliferation,migration and invasion of gastric cancer cells through targeted regulation of miR-5195-3p

目的探讨长链非编码RNA CBR3-AS1(lncRNA CBR3-AS1)是否通过靶向调控miR-5195-3p的表达从而促进胃癌细胞增殖、迁移及侵袭。方法体外培养正常胃上皮细胞GES-1与胃癌细胞系HGC-27、NCI-N87、AGS,采用实时荧光定量聚合酶链反应(qRT-PCR)检测CBR3-AS1、miR-5195-3p的表达量;分别将si-NC、si-CBR3-AS1、si-CBR3-AS1与anti-miR-NC、si-CBR3-AS1与anti-miR-5195-3p转染至AGS细胞;甲基噻唑基四唑(MTT)检测细胞增殖;Transwell小室实验检测细胞迁移及侵袭;双荧光素酶报告实验验证CBR3-AS1、miR-5195-3p的靶向关系;蛋白免疫印迹法(Western blotting)检测上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)的表达量。结果与GES-1比较,胃癌细胞系HGC-27、NCI-N87、AGS中CBR3-AS1的表达水平显著升高(P<0.05);与si-NC组比较,si-CBR3-AS1组细胞活力显著降低(P<0.05),迁移细胞数和侵袭细胞数显著减少(P<0.05),E-cadherin蛋白水平显著升高(P<0.05),N-cadherin蛋白水平显著降低(P<0.05);双荧光素酶报告实验证实CBR3-AS1可靶向结合miR-5195-3p;与si-CBR3-AS1+anti-miR-NC组比较,si-CBR3-AS1+anti-miR-5195-3p组细胞活力显著升高(P<0.05),迁移细胞数和侵袭细胞数显著增多(P<0.05),E-cadherin蛋白水平显著降低(P<0.05),N-cadherin蛋白水平显著升高(P<0.05)。结论CBR3-AS1可能通过抑制miR-5195-3p的表达从而促进胃癌细胞增殖、迁移及侵袭。

Objective To investigate whether lncRNA CBR3-AS1 can promote the proliferation,migration and invasion of gastric cancer cells by targeting and regulating the expression of miR-5195-3p.Methods Normal gastric epithelial cells GES-1 and gastric cancer cell lines HGC-27,NCI-N87,AGS were cultured in vitro,and the expression levels of CBR3-AS1,miR-5195-3p were detected by qRT-PCR.Si-NC,si-CBR3-AS1,si-CBR3-AS1 and anti-miR-NC,si-CBR3-AS1 and anti-miR-5195-3p were transfected into AGS cells,respectively.MTT was used to detect the cell proliferation.Transwell cell test was used to detect the cell migration and invasion.The double luciferase report experiment verified the targeting relationship of CBR3-AS1,miR-5195-3p.Western blotting was used to detect the expression levels of E-cadherin and N-cadherin.Results Compared with GES-1,the expression levels of CBR3-AS1 in gastric cancer cell lines HGC-27,NCI-N87,and AGS were significantly increased(P<0.05).Compared with the si-NC group,the cell viability of the si-CBR3-AS1 group was significantly reduced(P<0.05),the number of migrating cells and invasive cells was significantly reduced(P<0.05),and the level of E-cadherin protein was significantly increased(P<0.05),the level of N-cadherin protein was significantly reduced(P<0.05).The double luciferase report experiment confirmed that CBR3-AS1 could target and bind to miR-5195-3p.Compared with the si-CBR3-AS1+anti-miR-NC group,the cell viability of the si-CBR3-AS1+anti-miR-5195-3p group was increased significantly(P<0.05),and the number of migrating cells and invasive cells was increased significantly(P<0.05),the level of E-cadherin protein was significantly reduced(P<0.05),the level of N-cadherin protein was significantly increased(P<0.05).Conclusion CBR3-AS1 may promote gastric cancer cell proliferation,migration and invasion by inhibiting the expression of miR-5195-3p.