二氯乙酸盐对宫颈癌细胞分裂增殖的影响及机制研究

Effect of dichloroacetate on the division and proliferation of cervical cancer cells and its mechanism

目的研究二氯乙酸盐(DCA)对宫颈癌细胞分裂增殖的影响及机制。方法人宫颈癌HeLa细胞分为实验组和对照组,实验组细胞以终浓度为1,5,10,20,50,100 mmol·L^(-1)二氯乙酸盐处理,对照组细胞则以等量生理盐水处理。用噻唑蓝(MTT)法及克隆形成实验检测HeLa细胞增殖情况;用流式细胞术检测HeLa细胞周期及凋亡情况;用蛋白免疫印迹法检测HeLa细胞B淋巴细胞瘤-2(Bcl-2)的表达。结果处理24 h后,HeLa细胞的相对生存率随着二氯乙酸盐作用浓度的升高逐渐降低(P <0. 05)。对照组及20,50 mmol·L^(-1)实验组HeLa细胞克隆形成率分别为(125. 17±8. 43)%,(58. 53±5. 36)%,(40. 53±3. 57)%;实验组HeLa细胞克隆形成率较对照组降低(P <0. 05)。干预24 h后,20,50 mmol·L^(-1)实验组HeLa细胞G_0/G_1、G_2/M期细胞比例高于对照组,S期细胞比例低于对照组(P <0. 05)。对照组及20,50 mmol·L^(-1)实验组HeLa细胞凋亡率分别为(3. 29±0. 36)%,(16. 58±3. 26)%,(22. 23±2. 12)%,实验组细胞凋亡率明显高于对照组(P <0. 05)。与对照组比较,实验组细胞Bcl-2蛋白相对表达量降低(P <0. 05)。结论二氯乙酸盐可显著抑制人宫颈癌HeLa细胞增殖,其作用机制可能与诱导细胞凋亡的发生有关。

Objective To explore the effect of dichloroacetate（ DCA）on the division and proliferation of cervical cancer cells and its mechanism. Methods Human cervical cancer HeLa cells were divided into test group and control group,the cells in test group were treated with final concentration of 1,5,10,20,50 and 100 mmol · L^-1 DCA,the cells in control group were treated with equal amount of normal saline.The HeLa cells proliferation were detected by methyl thiazolyl tetrazolium（ MTT） and clone formation assay; the HeLa cells apoptosis and cell cycle were detected by flow cytometry. The expression of B-cell lymphoma-2（ Bcl-2） in HeLa cells were detected by Western Blot（ WB）.Results After treatment for 24 h,the relative survival rate of HeLa cells decreased gradually with the increase of DCA concentration（ P〈0. 05）.The clone formation rates of HeLa cells in control group and 20,50 mmol·L^-1 test group were（ 125. 17 ± 8. 43） %,（ 58. 53 ± 5. 36） %,（ 40. 53 ± 3. 57） %; the clone formation rates of HeLa cells in test group were lower than that of control group（ P〈0. 05）. After intervention for 24 h,the proportion of G0/G1 and G2/M phase cells in 20,50 mmol·L^-1 test group were higher than those in control group,and the proportion of S phase cells were lower than that of control group（ P〈0. 05）. The apoptosis rates of HeLa cells in control group and 20,50 mmol · L^-1 test group were（ 3. 29 ± 0. 36） %,（ 16. 58 ± 3. 26） %,（ 22. 23 ± 2. 12） %,the apoptosis rates of test group were significantly higher than that of control group（ P〈0. 05）.Compared with control group,the relative expression of Bcl-2 protein in test group decreased（ P〈0. 05）.Conclusion DCA can significantly inhibit the proliferation of human cervical cancer HeLa cells,which may be related to the induction of apoptosis.