LncRNA-SNHG12通过miR-409-3p/ZEB1轴调节宫颈癌细胞的增殖、凋亡和上皮间质转化

LncRNA-SNHG12 regulates proliferation,apoptosis and epithelial-mesenchymal transition of cervical cancer cells via the miR-409-3p/ZEB1 axis

目的探索长链非编码RNA(lnc RNA)SNHG12通过微小RNA(miR)-409-3p/锌指E盒结合的同源盒蛋白1(ZEB1)轴对宫颈癌细胞增殖、凋亡和上皮间质转化的影响。方法选取2021年2月至2022年2月于河南省南阳市中心医院诊治的50例宫颈癌患者组织及癌旁组织标本、正常宫颈上皮细胞以及宫颈癌细胞系He La、Si Ha、Ca Ski,检测组织及细胞中lnc RNA SNHG12与miR-409-3p的表达水平。取宫颈癌细胞系He La分为sh-NC组(转染阴性对照载体)、shSNHG12组(转染SNHG12敲低载体)、mimics NC组(转染模拟物阴性对照)、miR-409-3p mimics组(转染miR-409-3p模拟物)、sh-NC+inhibitor NC组(共转染sh-NC与抑制物阴性对照)、sh-NC+miR-409-3p inhibitor组(共转染sh-NC与miR-409-3p抑制物)、sh-SNHG12+inhibitor NC组(共转染sh-SNHG12与inhibitor NC)、sh-SNHG12+miR-409-3p inhibitor组(共转染sh-SNHG12与miR-409-3p inhibitor);未转染细胞为对照组。依次采用q RT-PCR、双荧光素酶实验检测SNHG12与miR-409-3p表达水平及两者的靶向关系;MTT法、流式细胞仪检测细胞增殖及凋亡率;Western blot检测神经钙黏蛋白(N-cadherin)、上皮钙黏蛋白(E-cadherin)、ZEB1、Snail和波形蛋白(vimentin)的表达水平。结果SNHG12与miR-409-3p存在靶向关系,与sh-NC组、对照组相比,shSNHG12组细胞凋亡率、E-cadherin蛋白表达显著增加,细胞增殖率、SNHG12、ZEB1、N-cadherin、Snail、vimentin表达显著降低(P<0.05);与mimics NC组、对照组相比,miR-409-3p mimics组细胞凋亡率、miR-409-3p、E-cadherin蛋白表达显著增加,细胞增殖率、ZEB1、N-cadherin、Snail、vimentin表达显著降低(P<0.05);miR-409-3p inhibitor可逆转敲低SNHG12对He La细胞发挥的上述作用(P<0.05)。结论敲低SNHG12表达可通过靶向负调控miR-409-3p/ZEB1轴,减少He La细胞增殖以及上皮间质转化,促进细胞凋亡。

Objective To explore the impacts of long non-coding RNA(lncRNA)SNHG12 on the proliferation,apoptosis and epithelial-mesenchymal transition of cervical cancer cells through microRNA(miR)-409-3p/zinc finger E-box-binding homeobox protein 1(ZEB1)axis.Methods Tissue and paracancer tissue samples of 50 patients with cervical cancer treated in our hospital from February 2021 to February 2022 were selected.The expression levels of LncRNA SNHG12 and miR-409-3p in normal cervical epithelial cells,cervical cancer cell lines HeLa,SiHa and CaSki,cancer tissue and paracancer tissue were determined.The cervical cancer cell lines HeLa were grouped into sh-NC group(transfected with negative control vector),sh-SNHG12 group(transfected with SNHG12 knockdown vector),mimics NC group(transfected with mimic negative control),miR-409-3p mimics group(transfected with miR-409-3p mimic),sh-NC+inhibitor NC group(co-transfected with sh-NC and inhibitor negative control),sh-NC+miR-409-3p inhibitor group(co-transfected with sh-NC and miR-409-3p inhibitor),sh-SNHG12+inhibitor NC group(co-transfected with sh-SNHG12 and inhibitor NC),and sh-SNHG12+miR-409-3p inhibitor group(co-transfected with sh-SNHG12 and miR-409-3p inhibitor),while untransfected cells were the control group.The expression levels of SNHG12 and miR-409-3p and their targeting relationship were detected by qRT-PCR and dual luciferase experiments respectively;the cell proliferation and apoptosis rate were detected by MTT method and flow cytometry;and the expression levels of N-cadherin,E-cadherin,ZEB1,Snail and vimentin were measured by Western blot.Results SNHG12 had a targeting relationship with miR-409-3p.Compared with the sh-NC group and the control group,the apoptosis rate and E-cadherin protein expression in the sh-SNHG12 group were notably increased,while the cell proliferation rate,and the SNHG12,ZEB1,N-cadherin,Snail and vimentin expression were notably decreased(P<0.05).Compared with the mimics NC group and the control group,the apoptosis rate,miR-409-3p and E-cadherin protein expressions in the miR-409-3p mimics group were notably increased,while the cell proliferation rate,and the ZEB1,N-cadherin,Snail and vimentin expressions were notably decreased(P<0.05).MiR-409-3p inhibitor was able to reverse the above effects of knockdown of SNHG12 on HeLa cells(P<0.05).Conclusions Knockdown of SNHG12 expression can reduce HeLa cell proliferation and epithelial-mesenchymal transition and promote apoptosis by targeting and negatively regulating the miR-409-3p/ZEB1 axis.