Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (27): 5062-5066.doi: 10.3969/j.issn.1673-8225.2010.27.028

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Anti-proliferation effect of Ponicidin on leukemia HL-60 cells

Liu Xiao-dan1, Liu Wen-da2, Wang Chun-zhi1, Xu Yan1, Lin Dong-jun1, Liu Pei-qing3, Huang He-qing3, Wu Chuan-bin3, Xiao Ruo-zhi1,Huang Ren-wei1, Liu Jia-jun1   

  1. 1 Department of Hematology, 2 Department of Transfusion, Third Hospital of Sun Yat-sen University, Guangzhou  510630, Guangdong Province, China; 3 Laboratory of Pharmacology and Toxicology, School of Pharmaceutical Science, Sun Yat-sen University, Guangzhou  510080, Guangdong Province, China
  • Online:2010-07-02 Published:2010-07-02
  • Contact: Liu Jia-jun, Associate professor, Master’s supervisor, Department of Hematology, Third Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China jiajun.l@163.com
  • About author:Liu Xiao-dan★, Studying for master’s degree, Department of Hematology, Third Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China lenovo381@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30772782*; the Outstanding Talent Support Project of Ministry of Education of China, No. NCET-0721*

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Abstract:

BACKGROUND: Recent studies have shown that Ponicidin (PON) plays a significant role in anti-cancer effect via inducing apoptosis in various human cancer cell lines. At present, few reports have addressed the PON effects on leukemia cell lines HL-60 cells.
OBJECTIVE: To investigate the anti-proliferation effect of PON on leukemic HL-60 cells in vitro and its mechanisms of action.
METHODS: HL-60 cells in culture medium in vitro were given different concentrations of PON (10-50 μmol/L) for 24, 48 and 72 hours. The inhibitory rates were measured by MTT assay, and cell cycle was detected by flow cytometry. Expressions of Caspase-3 and poly(ADP-ribose) polymerase (PARP) as well as cell cycle modulator P16 and P21 were detected by Western blot assay.
RESULTS AND CONCLUSION: PON could remarkably inhibit the growth of HL-60 cells and cause apoptosis; the suppression was in time- and dose-dependent manners. Flow cytometry detection revealed that the cells were mainly arrested in G0/G1 phase and the subG1 cells were also found clearly in the histogram following 50 μmol/L PON treatment. Western blot results showed cleavage of the Caspase-3 zymogen protein (Mr32 000), with the appearance of its Mr 20 000 subunit, and a cleaved Mr 89 000 fragment of PARP after the cells were treated by different concentrations of PON. Western blot analysis also revealed that P21 and P16 expression was gradually up-regulated following 50 μmol/L PON treatment. Results have indicated that PON demonstrates in vitro anti-leukemia effect in HL-60 cells mainly associated with cell cycle arrest and apoptosis. Up-regulation of the expression of P16 and P21 as well as activation of Caspase-3 may be one of its most important mechanisms of inducing HL-60 cell inhibition in G0/G1 phase or cell apoptosis.

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