Down-regulation of ANXA7 decreases metastatic potential of human hepatocellular carcinoma cells in vitro

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• 作者：Mohammed Mohammed Ibrahim^a ; Ming-Zhong Sun^b ; Yuhong Huang^a ; Mao Jun^a ; Yanling Jin^a ; Du Yue^a ; Wang Jiasheng^b ; Jun Zhang^a ; Asma Saleem Qazi^a ; Ken Sagoe^c ; Jianwu Tang^a ; ^{jwtang_53@yahoo.cn}
• 关键词：ANXA7 (Annexin A7) down-regulation ; Human hepatocellular carcinoma ; HepG2 cell migration and invasion
• 刊名：Biomedicine and Pharmacotherapy
• 出版年：2013
• 出版时间：May, 2013
• 年：2013
• 卷：67
• 期：4
• 页码：285-291
• 全文大小：1508 K

文摘

We report for the first time the influence of ANXA7 gene on human hepatocellular carcinoma cells (HCC). We down-regulated ANXA7 in human HCC cell line (HepG2) using siRNA method. By Western Blot analysis, we confirmed about 70 % down-regulation of the gene in the shRNA-ANXA7 transfected cells (shRNA-ANXA7-HepG2) compared to the non-specific sequence shRNA transfected cells (control-shRNA-HepG2) and the un-manipulated-HepG2 cells. We used CCK-8 cell proliferation kit and observed about 65 % reduction in the shRNA-ANXA7-HepG2 cells where the two controls exhibited comparable cell proliferation rates. Also, by using PI staining followed by flow cytometry, we noticed a cell cycle arrest at G₀/G₁ with more than one fold reduction of shRNA-ANXA7-HepG2 cell population in the S-phase of the cell cycle. Also of particular note was a significant aneuploidy in the controls compared to zero aneuploidy in the ANXA7 down-regulated cells. Migration of the cells was detected using Boyden's transwell chamber and scratch wound healing assay which showed 50 % and 30 % respective reductions in shRNA-ANXA7-HepG2 cells migration. Furthermore, the control-shRNA-HepG2 cells and the un-manipulated-HepG2 cells invaded through the ECM-coated transwell plates two times more than the shRNA-ANXA7-HepG2 cells. We have found ANXA7 to be functioning like a tumour promoter in HepG2 human hepatocellular carcinoma cells and could have a potential as a therapeutic window into the management of liver cancer.