关键词: 3-溴丙酮酸, HCT116, 己糖激酶Ⅱ, 6-磷酸葡萄糖

Abstract: Abstract:Objective To study the inhibitory effect of 3-bromopyruvate(3-BP) on proliferation of colon carcinoma HCT116 cells  and to explore its mechanism,and to provide  evidence for the clinical application of 3-BP.Methods HCT116 cells were cultivated in vitro and divided into control group and different concentrations of 3-BP groups(25,50,75,and 100 mg/L).The hexokinaseⅡ(HKⅡ) protein expression levels in various groups were determined by Western blotting method;the concentrations of glucose-6-phosphatase (G-6-P) were determined by G-6-P a
ssay kit;the proliferation activities of the cells were observed by MTT method;the cell cycle was measured by flow cytometry. Results Compared with control group,the HKⅡ protein level in 25 mg/L 3-BP group did not change significantly（P>0.05）;compared with control group and 25 mg/L 3-BP group,the HKⅡprotein levels in 50,75, and 100 mg/L 3-BP groups were decreased significantly（P<0.01）.Compared with control group,the concentration of G-6-P in culture medium in 25 mg/L 3-BP group did  not change significantly（P>0.05）;compared with control group and 25 mg/L 3-BP group,the concentration of G-6-P in culture medium in 50,75,and 100 mg/L 3-BP groups  were decreased significantly（P<0.01）.The MTT results showed that compared with control group,the survival rate in 25 mg/L 3-BP group did  not change significantly(P>0.05);compared with control group and 25 mg/L 3-BP group,the survival rates
 in 50,75,and 100 mg/L 3-BP groups were decreased markedly(P<0.05).The flow cytometry analysis results showed that compared with control group,the percentages of the cells at G1 phase and S phase  in 25 mg/L 3-BP group did  not  change significantly（P>0.05）;compared with  control group and  25 mg/L 3-BP group,the percentages of the cells at G1 phase  in 50,75 and 100 mg/L 3-BP groups were increased significantly(P<0.01)  and the percentages of the cells at S phase  were decreased(P<0.05).The cell cycle was arrested at G1 stage.Conclusion 3-BP could inhibit the proliferation of  HCT116 cells  significantly,which  may be related to the decreasing of HKⅡ protein expression and  use of glucose.

Key words: 3-bromopyruvate, HCT116, hexokinaseⅡ, glucose-6-phosphate