miR-19a在肝癌组织中高表达对肝癌HepG2细胞上皮-间质样转化的影响

Effects of high expression of miR 19a in hepatocellular carcinoma tissues on the epithelial mesenchymal transition in HepG2 cells

目的探讨微小RNA(microRNA,miRNA,miR)-19a在肝癌组织中的表达及miR-19a对肝癌HepG2细胞上皮-间质样转化(EMT)转化的影响。方法应用实时荧光定量聚合酶链式反应(quantificational real-time polymerase chainreaction,qRT-PCR)方法检测了25例肝癌及其相应癌旁组织中miR-19a的表达。筛选miR-19a低表达的肝癌细胞并将miR-19a模拟物(mimic)或其阴性对照(miR-negative control,miR-NC)转染至HepG2细胞后,应用qRT-PCR检测HepG2细胞中miR-19a的表达水平,MTT和Transwell小室法分别检测细胞的增殖能力和迁移能力。应用实时荧光定量PCR和蛋白质印迹法分别检测转染miR-19a-mimic后,HepG2细胞中E-cad、N-cad和Vimentin mRNA及其蛋白的表达水平。结果肝癌组织中miR-19a的表达水平明显高于癌旁组织(P<0.01)。miR-19a-mimic转染后,HepG2细胞中miR-19a的表达水平明显高于阴性对照组(P<0.01),细胞的增殖能力及迁移能力均明显高于阴性对照组(P<0.01)。miR-19a-mimic转染后,HepG2细胞中E-cad mRNA及蛋白的表达水平明显低于阴性对照组(P<0.01),N-cad和Vimentin mRNA及蛋白的表达水平明显高于阴性对照组(P<0.01)。结论 miR-19a在肝癌组织中呈高表达状态,促进肝癌HepG2细胞的增殖及迁移能力,诱导肝癌HepG2细胞的EMT。

Objective To investigate the expression of microRNA（ microRNA,miRNA,miR） 19 a in hepatocellular carcinoma tissues and the effects of miR-19 a on epithelial-mesenchymal transition（ EMT） in hepatocellular cell line-HepG2.Methods Quantificational real time polymerase chainreaction（ qRT PCR） was used to measure the expression levels of miR-19 a in hepatocellular carcinoma tissues and corresponding para-carcinoma tissues in 25 patients with hepatic carcinoma.The MiR 19 a mimic or negative control of miR 19 a（ miR NC） were transfected into HepG2 with low expressions of miR 19 a.The expression levels of miR 19 a in HepG2 cells were detected by qRT PCR,moreover MTT and Transwell small chamber method were used to measure the abilities of proliferation and migration of the cells. In addition,the expression levels of Ecad,N-cad,Vimentin mRNA and proteins were detected by qRT PCR and Western Blot after miR 19 a mimic was transfected into HepG2 cells. Results The expression levels of miR-19 a in hepatocellular carcinoma tissues were significantly higher than those in paracancerous tissues（ P 〈0. 01）. After miR-19 a-mimic was transfected into HepG2 cells,the expression levels of miR-19 a were significantly higher than those in negative control group（ P 〈0. 01）,and the proliferation and migration abilities of cells were significantly higher than those in negative control group（ P 〈0. 01）. Moreover the expression levels of E-cad mRNA and proteins were significantly lower than those in negative control group（ P 〈0. 01）,however the expression levels of N cad,Vimentin mRNA and proteins in HepG2 cells were significantly higher than those in negative control group（ P〈 0. 01）. Conclusion The high expression levels of MiR 19 a exist in hepatocellular carcinoma tissues,which can promote the proliferation and migration ablities of HepG2 cells and induce the EMT of HepG2 cells.