HMG20A对肝癌细胞体外增殖与迁移的影响及其机制

Effect and mechanism of HMG20A on the proliferation and migration of hepatocellular carcinoma cells

目的研究HMG20A在肝细胞癌(hepatocellular carcinoma,HCC)发生发展和转移中的功能与机制。方法在不同转移能力和遗传背景的肝癌细胞Huh7和HCCLM3中分别构建HMG20A过表达和敲低稳定株,通过实时定量PCR(real time quantitative PCR,qPCR)验证过表达和敲低该基因的效果。用CCK8试剂盒检测HMG20A对肝癌细胞增殖能力的影响,利用Transwell小室分析HMG20A调控肝癌细胞转移的能力。借助Western blot和qPCR分析HMG20A调控肝癌增殖和转移的机制。结果体外实验表明,HMG20A能够促进肝癌细胞的体外增殖和迁移,显著上调促分裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路中p38(p38 MAPK)、细胞外调节蛋白激酶(extracellular regulated protein kinase,ERK)的活性和表达水平,同时上皮间充质转化(epithelial-mesenchymal transition,EMT)的标志物Vimentin、抗平滑肌抗体(anti-smooth muscle antibody,alpha-SMA)、N-cadherin受到显著的正调控,E-cadherin受到显著负调控。结论 HMG20A可能通过促进EMT进程和MAPK通路促进肝癌细胞的体外增殖与迁移。

Objective To study the mechanism of HMG20A in the development and metastasis of hepatocellular carcinoma（HCC）. Methods We constructed HMG20A overexpression and knock-down stable cell lines in Huh7 and HCCLM3 which had different metastatic abilities and genetic backgrounds.The over expression and knock down effects of HMG20A were analyzed by real time quantitative PCR （qPCR）.The effect of HMG20A on the proliferation of HCC cell lines was detected by cell counting kit-8 assay.The Transwell analysis was proceeded to assess the effect of HMG20A on metastasis of HCC cells.We investigated the mechanism of HMG20A in proliferation and metastasis of HCC cells using qPCR analysis and Western blot. Results The in vitro experiments suggested that HMG20A could promote the proliferation and metastasis of HCC.It also showed that HMG20A could upregulate the activity and expression levels of mitogen-activated protein kinases （MAPKs） such as p38 and extracellular regulated protein kinase （ERK）.Futhermore,it could regulate the expression levels of the biomarkers of epithelial-mesenchymal transition （EMT） such as Vimentin,anti-smooth muscle antibody （alpha-SMA） and N-cadherin positively and E-cadherin negatively. Conclusions HMG20A may promote the proliferation and migration of HCC cells in vitro by promoting the EMT process and MAPK pathway.