Inhibitory effect of metformin on proliferation of megakaryocytic leukemia cell line Dami and its mechanismChinese Full Text
LI Chang-ling;LIN Di;XING Si-ning;ZHAO Song;CHEN Hui-peng;ZHOU Fan;MA Dong-chu;Department of Experimental Medicine,General Hospital,Shenyang Military Area Command;Institute of Biological Engineering,Academy of Military Medical Sciences;Department of Hematology,General Hospital,Shenyang Military Area Command;
Abstract: Objective To study the effect of metformin on the growth of megakaryocytic leukemia cell line Dami and to explore the molecular mechanisms of the inhibitory effect of metformin on the proliferation of Dami. Methods The Dami cells were cultured and divided into control and 1,2,4,8,16and 32mmol·L-1 metformin groups.Then MTT test was performed to detect the inhitory rate of proliferation of Dami cells after treated with different concentrations of metformin.Flow cytometry was used to examine the distribution of cell cycle,and Western blotting was carried out to analyze the expressions of Cdc2and CylinB1and the phosphorylation of Cdc2. Results The MTT results showed that compared with control group,the inhibitory rates of proliferation of the Dami cells in 32 mmol·L-1 metformin groups at 0,24,48,72and 96h（35.1% ±2.3%,49.7% ±5.1%, 78.85±0.9%,79.1%±3.0%%,and 85.2% ±3.2%）were significantly increased（P＜0.01）,Furthermore, after metformin treatment for 72h,the inhibitory rates of proliferation of the Dami cells in 1,2,4,8,16and 32mmol·L-1 metformin groups were（33.8±0.3）%,（51.9±0.2）%,（59.4±1.6）%,（65.5±2.0）%,（75.5±0.9）%,and（79.1±3.0）%,respectively.Metformin inhibited the growth of Dami cells in a time-and dose-dependent manner.The flow cytometry results results revealed that compared with control group,the percentages of Dami cells in G2/M phase in 1,2and 4mmol·L-1 metformin groups were increased from（26.0± 0.5）%to（38.5±1.5）%,（48.4±1.1）%,and（58.2±2.7）%;there was significant difference in the percentages of Dami cells in G2/M phase between control group and 4mmol·L-1 metformin group（P＜0.01）. Western blotting analysis showed that compared with control group,the expressions of Cdc2and CyclinB were evidently reduced,the phosophorylation of Cdc2at Tyr15was up-regulated,and the phosphorylation at Thr161 was down-regulated.Conclusion Metformin can inhibit the growth of Dami cells and induce G2/M arrest,and its mechanism may be related to inhibiting the activation of Cdc2/CyclinB1complex.
- DOI：
10.13481/j.1671-587x.20140314
- Series：
- Subject：
- Classification Code：
R733.7
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