Induction of H2AX phosphorylation in tumor cells by gossypol acetic acid is mediated by phosphatidylinositol 3-kinase (PI3K) family

Cancer Cell International

Shang-Biao Li¹  Chao Jiang¹  Shu-Yan Pei¹  Chen-Jing Wang¹  Jian-Xiu Ma¹  Lei Song¹  Jin Zhao¹  Zhong Guo¹ 

[1] Medical College of Northwest University for Nationalities, Lanzhou 730030, PR China

关键词: ATR;    ATM;    DNA-PK;    H2AX phosphorylation;    Gossypol acetic acid;   

Others  :  1121603
DOI  :  10.1186/s12935-014-0141-5

 received in 2014-06-26, accepted in 2014-12-04,  发布年份 2014

【 摘 要 】

Background

H2AX is phosphorylated (γH2AX) by members of the phosphatidylinositol 3-kinase (PI3K) family, including Ataxia telangiectasia-mutated (ATM), ATM- and Rad3-related (ATR) and DNA-PK in response to DNA damage. Our study shows that gossypol acetic acid (GAA) alone can induce γH2AX in Human mucoepidermoid carcinoma cell line (MEC-1) in vitro. Thus, we further examined the possible mechanisms of GAA to induce γH2AX in tumor cells.

Materials and methods

The PI3K inhibitors caffeine and wortmannin were used in an effort to identify the kinase(s) responsible for GAA -induced γH2AX in MEC-1 cells. DNA dependent protein kinase (DNA-PK) - proficient and –deficient cells, human glioma cell lines M059K and M059J, were also used to evaluate the kinases responsible for GAA induced H2AX phosphorylation. γH2AX expression was detected by immunofluorescent microscopy. Flow cytometry assay was used to assay γH2AX and cell cycle.

Results

GAA induced H2AX phosphorylation in a cell cycle-dependent manner and a significant G0/G1 phase arrest in MEC-1 cells was shown. Caffeine and wortmannin significantly inhibited GAA-induced H2AX phosphorylation in MEC-1 cells. GAA induced H2AX phosphorylation in M059K, but not in M059J. Taken together, these data suggested that GAA treatment alone could induce H2AX phosphorylation in a cell cycle dependent manner in MEC-1 and M059K, but not in M059J cells. A significant G0/G1 phase arrest was shown in MEC-1.

Conclusions

The member of PI3K family, DNA-PK, ATM and ATR are involved in the H2AX phosphorylation of MEC-1 cells.

【 授权许可】

   
2014 Guo et al.; licensee BioMed Central Ltd.

【 图 表 】

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