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
阿木提江·马合木提 1买热帕提·艾尔凯西 2郑坚江 1迪里夏提·阿里木 1陈雄1
作者信息
- 1. 新疆维吾尔自治区人民医院肝胆胰诊疗中心胰腺科 新疆乌鲁木齐 830001
- 2. 新疆医科大学第八附属医院影像科 新疆乌鲁木齐 830001
- 折叠
摘要
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BxPc-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,niR-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.
Abstract
Objective:To investigate the targeted regulation of high mobility group A2(HMGA2)expression by miR-194-5p,thereby inhibiting epithelial mesenchymal transformation(EMT)induced by transforming growth factor(TGF)-β 1 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 for 72 h,what's more,miR-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 miR-194-5p,Western blot was to detect protein expression of HMGA2 and EMT markers(E-cadherin,Vimentin,and N-cadherin),Tran-swell 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 miR-194-5p,HMGA2,Vimentin,and N-cadherin were significantly higher,while the expression level of E-cadherin was significantly lower,the number of inva-sive cells was less,too(P＜0.05).Compared with the treatment group and Sh-NC group,the morphological changes of cells in the Sh-miR-194-5p group were significantly less,but still more than the control group,the expression levels of miR-194-5p,HMGA2,Vi-mentin,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 miR-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.
关键词
胰腺癌/上皮—间质转化/转化生长因子-β1/高迁移率族蛋白2/miR-194-5pKey words
Pancreatic cancer/Epithelial mesenchymal transition/Transforming growth factor-β1/High mobility group A2/MiR-194-5p引用本文复制引用
基金项目
新疆少数民族科技人才特殊培养计划科研项目(2021D03019)
出版年
2024
NETL
NSTL
万方数据