title: Down-regulation of ANXA7 decreases metastatic potential of human hepatocellular carcinoma cells in vitro.
abstract: 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.
authors.affiliation: Department of Pathology and Forensic Medicine, Dalian Medical University, 9 West Lvshun Southern Road, Dalian 116044, PR China.
authors.forename: Mohammed Mohammed
authors.initials: MM
authors.lastname: Ibrahim
authors.forename: Ming-Zhong
authors.initials: MZ
authors.lastname: Sun
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Huang
authors.forename: Mao
authors.initials: M
authors.lastname: Jun
authors.forename: Yanling
authors.initials: Y
authors.lastname: Jin
authors.forename: Du
authors.initials: D
authors.lastname: Yue
authors.forename: Wang
authors.initials: W
authors.lastname: Jiasheng
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Asma Saleem
authors.initials: AS
authors.lastname: Qazi
authors.forename: Ken
authors.initials: K
authors.lastname: Sagoe
authors.forename: Jianwu
authors.initials: J
authors.lastname: Tang
mesh_terms: D000782:Aneuploidy; D017310:Annexin A7 / Q000235:genetics*; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D059447:Cell Cycle Checkpoints / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D015536:Down-Regulation; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D009361:Neoplasm Invasiveness / Q000235:genetics; D009362:Neoplasm Metastasis; D034741:RNA, Small Interfering / Q000008:administration & dosage
chemical_list: D017310:Annexin A7; D034741:RNA, Small Interfering