沉默TGFA抑制肝癌细胞增殖的研究

Knockdown of TGFA Inhibits the Growth of Hepatocellular Carcinoma

目的探索siRNA沉默TGFA后对肝癌细胞增殖的影响。方法将TGFA-siRNA转染至L02和HepG2细胞中,Real-timePCR检测TGFAmRNA表达效率,Western blot检测TGFA的蛋白表达水平;MTT和BrdU法检测沉默TGFA后L02和HepG2细胞增殖的变化,Western blot检测TGFA/EGFR通路的活性;Real-time PCR检测肝癌组织中TGFA与EGFR的mRNA表达并分析其相关性。结果在L02和HepG2细胞中,TGFA-siRNA可下调TGFA的表达,对肝癌细胞增殖有抑制作用而不影响正常肝细胞增殖,并可抑制TGFA/EGFR通路活性。肝癌组织标本中TGFA与EGFR的mRNA均高表达且呈正相关。结论沉默TGFA基因可抑制肝癌细胞HepG2的增殖。

Objective To investigate the effects of knockdown of TGFA on the proliferation of hepatocellular carcinoma cells. Methods We selected TGFA as the candidate target gene. TGFA-si RNA was transfected into L02 and HepG2 cells. The expression efficiency of TGFA m RNA was detected by Real-time PCR, and the expression level of TGFA protein was detected by Western blot. Then MTT and Brd U methods were used to detect the effects of down-regulation of TGFA-si RNA on proliferation of L02 and HepG2 cells. Western blot assay was applied to detect the TGFA/EGFR pathway activity. TGFA over expression was achieved to verify the correlation between TGFA and EGFR expression in hepatocellular carcinoma. Results TGFA-si RNA down-regulated the expression of TGFA, inhibited proliferation of human hepatoma cells but had no obvious effects on normal cell line L02. It also inhibited the activity of TGFA/EGFR pathways. The m RNA expression levels of TGFA and EGFR were specifically high in the hepatocellular carcinoma tissue and were positively correlated. Conclusion Knockdown of TGFA inhibited the proliferation of HepG2 hepatocellular carcinoma cells.