MiR-194-5p靶向调控HMGA2抑制TGF-β1诱导的胰腺癌EMT的作用

MiR-194-5p Targeted Regulation of HMGA2 to Inhibit EMT Induced by TGF-β1 in Pancreatic Cancer

目的:研究miR-194-5p靶向调控高迁移率族蛋白2(HMGA2)表达,进而抑制转化生长因子(TGF)-β1诱导的胰腺癌细胞上皮-间质转化(EMT)的相关分子机制。方法:将人胰腺癌细胞系Bx Pc-3分为四组,分别为对照组、处理组、Sh-miR-194-5p组和Sh-NC组,除了对照组,其他三组均在体外采用TGF-β1诱导培养72 h,分别构建miR-194-5p沉默质粒(Sh-miR-194-5p)和对照质粒(Sh-NC)在TGF-β1诱导培养前2 h转染细胞。倒置显微镜观察细胞形态,qRT-PCR法检测miR-194-5p,Western blot法检测HMGA2以及EMT标志物(E-cadherin、Vimentin和N-cadherin)蛋白,Transwell实验检测细胞侵袭。结果:与对照组相比,处理组细胞形态为长梭形,出现EMT特征性形态;miR-194-5p、HMGA2、Vimentin和N-cadherin表达量明显升高,而E-cadherin表达量明显下降,侵袭细胞数目减少(P<0.05)。与处理组和Sh-NC组相比,Sh-miR-194-5p组细胞形态变化明显减少,但仍多于对照组,miR-194-5p、HMGA2、Vimentin和N-cadherin表达量明显下降,而E-cadherin表达量明显升高,侵袭细胞数目增多(P<0.05),与对照组相比差异仍有统计学意义(P<0.05)。结论:人胰腺癌细胞可高表达miR-194-5p,通过靶向调控HMGA2表达进而部分程度上抑制了TGF-β1诱导的细胞EMT变化,为疾病发生机制以及临床干预提供了新型靶点。

Objective:To investigate the targeted regulation of high mobility group A2(HMGA2)expression by mi R-194-5p,thereby inhibiting epithelial mesenchymal transformation(EMT)induced by transforming growth factor(TGF)-β1 in pancreatic cancer cells.Methods:Human pancreatic cancer cell line BxPc-3 was divided into four groups:control group,treatment group,Sh-miR-194-5p group and Sh-NC group.Except the control group,the other three groups were treated with TGF-β1 in vitro to induce and cultivate for72 h,what's more,mi R-194-5p silencing plasmid(Sh-miR-194-5p)and control plasmid(Sh-NC)were constructed to transfect cells 2 h before TGF-β1 induced culture.The morphological changes of cells were observed under inverted microscope,qRT-PCR was to detect mi R-194-5p,Western blot was to detect protein expression of HMGA2 and EMT markers(E-cadherin,Vimentin,and N-cadherin),Transwell assay was to detect the invasiveness of cells.Results:Compared with control group,the cell morphology of the treatment group changed to a long spindle shape,exhibiting characteristic morphological changes of EMT;the expression levels of mi R-194-5p,HMGA2,Vimentin,and N-cadherin were significantly higher,while the expression level of E-cadherin was significantly lower,the number of invasive cells was less,too(P<0.05).Compared with the treatment group and Sh-NC group,the morphological changes of cells in the Sh-mi R-194-5p group were significantly less,but still more than the control group,the expression levels of mi R-194-5p,HMGA2,Vimentin,and N-cadherin were significantly lower,but the expression level of E-cadherin was significantly more and number of invasive cells was more,too(P<0.05),the differences between the Sh-miR-194-5p group and the control group were still statistically significant(P<0.05).Conclusion:Human pancreatic cancer cells may overexpress mi R-194-5p,which partially inhibits cell EMT changes induced by TGF-β1 via targetly regulating HMGA2 expression.It can provide novel targets for disease pathogenesis and clinical intervention.