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The Effects of Specific Down-regulation of Grp78 by siRNA on Invasion and Metastasis of Hepatocellular Carcinoma
Rong-Jian Su*Δ, Zhen Li1Δ, Liu-Fang Cheng1, Jia Wei, Hong-Dan Li
Central Laboratory, Liaoning Medical College, Jinzhou 121001, China; 1Department of Gastroenterology, General Hospital of Chinese Liberation Army, Beijing 100583, China
Abstract: To investigate the roles of glucose-regulated protein 78 (Grp78) in invasion and metastasis of hepatocellular carcinoma (HCC)， we down-regulated the expression of Grp78 by small interfering RNA (siRNA) in human HCC cell lines BEL 7402， the invasion and metastasis behaviors of HCC in Grp78 knock-down cells was analyzed by Transwell assay and wound healing assay. Focal adhesion kinase (FAK) phosphorylation was analyzed by immunoprecipitation. The activity of RhoA was determined by GST-pulldown assay. The expression status of E-cadherin， N-cadherin and vimentin was examined by Western blot. We found that the invasion and metastasis capabilities of HCC in Grp78 knock-down cells were significantly decreased compared with parental cells. Phosphorylation of FAK was significantly inhibited and RhoA activity was promoted in Grp78 knock-down cells. The expression levels of N-cadherin， vimentin were significantly lower， while E-cadherin was higher in Grp78 knock-down cells than in normal cells. We concluded that Grp78 silencing may inhibit the invasion and metastasis of HCC， These data suggest that Grp78 may be a potential target for HCC gene therapy.
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