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Apoptosis Induced by 3-Bromopyruvic Acid on Human Osteosarcoma Cell Line MG63

Author: LiKeQian
Tutor: ShaoZengWu
School: Huazhong University of Science and Technology
Course: Surgery
Keywords: 3-bromopyruvic acid hexokinase cisplatin apoptosis MG63 cell
CLC: R738
Type: Master's thesis
Year: 2011
Downloads: 3
Quote: 0
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Abstract


Objective: The aim of this study is to investigate the effect of 3-bromopyruvic acid inducing apoptosis on osteosarcoma cell line MG63 in vitro, to determine if there is a synergistic effect of 3-bromopyruvic acid(3-BrPA) combined with cisplatin(DDP) and to find out the possible mechanism of apoptosis induced by 3-bromopyruvic acid.Methods: The osteosarcoma cell line MG-63 were cultured regularly in the incubation box of 37℃and 5%CO2 , supplemented with DMEM/F12 containing 10% fetal bovine serum. After treated with different concentration of drugs, apoptotic cells morphology was observed at inverted phase contrast microscope. Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the effects on cell proliferation; Flow Cytometry(FCM) was used to analyze the effects on cell apoptosis. Real-time PCR technique was used to detect the expression level of hexokinase2 mRNA in osteosarcoma cells. Statistic analysis was processed by SPSS12.0 software.Results: The result of MTT and FCM showed that 3-bromopyruvic acid can inhibit MG63 cells and induce apoptosis. The anti-proliferation rate rised with the increase of drugs dose and time. Combination treatment of low-dose 3-bromopyruvic acid and cisplatin can significantly inhibited the cell growth compared with low-dose 3-bromopyruvic acid or cisplatin respectively (P<0.05). The biggest apoptosis rate of Combination treatment of low-dose 3-bromopyruvic acid and cisplatin was 73.18%. The hexokinase2 mRNA expression level in the experiment group was significantly lower than the control group in the real-time PCR experiment.Conclusions: 3-bromopyruvic acid inhibit MG63 cells and induce apoptosis in the dose- and time-dependent manner;Low-dose 3-bromopyruvic acid combining with cisplatin had a synergistic effect on inducing apoptosis in MG63 cells;The apoptosis induced by 3-bromopyruvic acid closely related to the downregulation of hexokinase2 mRNA.

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