Gli2基因沉默逆转肝癌细胞系SMMC-7721的上皮间质转化

摘要

Objective To explore the effect and mechanism of Gli2 on EMT and invasion in the hepatocellular car-cinoma cell line SMMC-7721.Methods shRNA of Gli2 and Negative control (NC) shRNA were constructed and transfected into SMMC-7721 cells.shRNA-Gli2 group,shRNA-NC group and blank group were set up .Transwell chambers assay , adhesion experiments were used to detect the ability of invasion ,homogeneous and heterogeneous cells intercellular adhesion of each group .Meanwhile, the qRT-PCR, Western blot were used to examine Gli2, E-cadherin ,N-cadherin and vimentin mRNA and protein expression .Results In different hepatocellular carcinoma cell lines and be along with the increasing ability of the invasion in hepatocellular carcinoma cell lines , Gli2 expres-sion was higher .Compared with the shRNA-NC group and blank control group ,the interfered group extensive cells invasion ability was inhibited ( P <0.05 ) , homogeneous cells intercellular adhesion increased and heterogeneous cells intercellular was opposite ( P <0.05 ) , meanwhile , the expression of E-cadherin was declined significantly ( P<0.05 ) ,the expression of N-cadherin , vimentin raised significantly ( P<0.05 ) .Conclusions Silencing Gli2 gene can reverse the EMT of the hepatocellular carcinoma cell line SMMC-7721 and inhibit cell invasion ,its mecha- nism may be related to the up-regulation of E-cadherin and the dow N-regulation of N-cadherin, vimentin.%目的：探讨胶质瘤相关癌基因同源蛋白2（Gli2）对肝癌细胞系SMMC-7721上皮间质转化（EMT）和侵袭能力的影响及机制。方法合成shRNA-Gli2、shRNA-NC病毒载体，实验设shRNA-Gli2组、shRNA-NC组和空白组，转染SMMC-7721细胞。用Transwell小室和细胞黏附实验观察细胞侵袭及黏附能力；用qRT-PCR和Western blot检测细胞间Gli2、E-cadherin、N-cadherin及vimentin基因和蛋白的表达。结果在不同肝癌细胞系中，随着肝癌细胞系侵袭能力的增强，Gli2的表达逐渐增高，shRNA-Gli2组与对照组相比侵袭能力降低，同种黏附能力增加，异种黏附能力降低（P＜0.05）。 shRNA-Gli2组与对照组相比，E-cadherin 表达明显增高而N-cadherin 和vimentin 表达明显降低（ P＜0.05）。结论沉默Gli2的表达能够促进肝癌细胞SMMC-7721 EMT的反转，并降低其侵袭能力，机制可能与E-cadherin的上调和N-cadherin、vimentin的下调有关。