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Lowered HGK expression inhibits cell invasion and adhesion in hepatocellular carcinoma cell line HepG2

World Journal of Gastroenterology 2010年第36期16卷 4541-4548页

作者： Su-Xia Han,Qing Zhu,Jin-Lu Ma,Jing Zhao,Xi Jia,Dan Zhang,Oncology Center of the First Aff iliated Hospital,College of Medicine,Xi’an Jiaotong University,Xi’an 710061,Shaanxi Province,ChinaChen Huang,Central Laboratory,College of Medicine,Xi’an Jiaotong University,Xi’an 710061,Shaanxi Province,China Oncology Center of the First Affiliated Hospital College of Medicine Xi'an Jiaotong University Xi'an 710061 Shaanxi Province China. hsummer22099@***

AIM:To investigate the effects of RNA interference tar-geting hepatocyte progenitor kinase-like kinase(HGK) in the invasion and adhesion of hepatocellular carcinoma(HCC) cell line ***:Three paired insert DNA fragments specif ic to HGK gene and one negative control DNA fragment were synthesized and inserted into RNAi-Ready pSIREN-RetroQ-ZsGreen *** blotting assay and real-time reverse transcriptase polymerase chain reaction(RT-PCR) were used to screen the vector with a highest inhibitory *** vector was used to generate recom-binant retrovirus specif ic to HGK.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2h-tetrazolium bromide(MTT) assay was used to examine cell growth;wound closure assay and cell adhesion assay were employed to investigate cell migration and adhesion respectively;and transwell assay and three-dimensional culture invasion assay were used to detect cell *** expressions of matrix metalloproteinase(MMP)-2,MMP-9 and nuclear factor(NF)-κB were detected by Western blotting ***:The real time RT-PCR and Western blotting assay showed that cells transfected with retrovirus me-diating RNAi targeting of HGK(RV-shHGK)-1 vector had the strongest inhibition of HGK protein,with an inhibi-tion rate of 76%,and this vector was used to generate recombinant retrovirus *** adhesion assay and MTT assay found that cell adhesion and growth of the cells infected with RV-shHGK-1 were significantly lower than those of the control cells(P < 0.05).Wound closure assay,transwell assay and three-dimensional culture invasion assay showed that the cell invasiveness was significantly less in HGK knockdown cells than in the control cells(P < 0.05).The expressions of MMP-2,MMP-9 and NF-κB were inhibited in HepG2 cells infected with ***:Down-regulation of HGK can obviously inhibit the migration and invasion of HepG2 cells in *** may be a new therapeutic target for treatment of HCC.

关键词： Hepatocellular carcinoma Hepatocyte pro-genitor kinase-like kinase RNA interference Invasion Metastasis

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