title: [Effects of Annexin A2 gene transcription down-regulation on human hepatoma cell biological behaviour].
abstract: OBJECTIVE
  To investigate the effects of Annexin A2 (ANXA2) deficiency on the malignant biological behaviour of hepatoma cells.
  
  
  METHODS
  The human hepatocellular carcinoma (HCC) cells lines MHCC97-H, HepG2, SMMC-7721, SMMC-7402 and L02 were evaluated. The expression and distribution of ANXA2 were analysed by western blotting, real-time PCR, immunofluorescence and immunohistochemistry.Cell cycle was assessed by flow cytometry and propidium iodide staining. Effects of ANXA2 silencing on invasion and migration potential were assessed by transwell assay and wound healing assay, respectively. Proliferative potential was assessed by CCK-8 kit in vitro and xenograft tumour-growth assay in vivo. The t-test, chi square test, rank sum test, q-test and F-test were used for statistical analyses.
  
  
  RESULTS
  The expression level of ANXA2 was markedly higher in the MHCC97-H cells with high metastasis potential than in the HepG2, SMMC-7721, SMMC-7402 and L02 cells. The efficiency of shRNA-mediated ANXA2 deficiency was more than 80%. Immunofluorescence analysis of the MHCC97-H cells indicated that ANXA2 expression was mainly localized to the cellular membrane and cytoplasm, with some nuclear localization. Down-regulation of ANXA2 led to S-phase arrest of HCC cells (q =8.001, P =0.002) and an inhibition of proliferation (q =17.140, P less than 0.01), migration (q =12.808, P less than 0.01) and invasion potential (q =9.069, P =0.002). Xenograft tumour-growth assay indicated that shRNA targeting of ANXA2 led to lower tumour weight (q =11.968, P < 0.001) and down-regulated ANXA2 expression (Z =2.530, P =0.011).
  
  
  CONCLUSIONS
  Down-regulation of Annexin A2 gene transcription effectively changes the biological behaviours of hepatoma cells, and may represent a potential target of HCC molecular therapies.
authors.affiliation: Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, China.
authors.forename: Haijian
authors.initials: H
authors.lastname: Zhang
authors.forename: Min
authors.initials: M
authors.lastname: Yao
authors.forename: Qi
authors.initials: Q
authors.lastname: Qian
authors.forename: Yun
authors.initials: Y
authors.lastname: Shi
authors.forename: Jingyuan
authors.initials: J
authors.lastname: Li
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.forename: Siye
authors.initials: S
authors.lastname: Wang
authors.forename: Dengfu
authors.initials: D
authors.lastname: Yao
mesh_terms: D000818:Animals; D017306:Annexin A2 / Q000235:genetics*; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D009368:Neoplasm Transplantation; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D014158:Transcription, Genetic
chemical_list: C472822:ANXA2 protein, human; D017306:Annexin A2; D034741:RNA, Small Interfering