二氯乙酸对膀胱癌细胞株T24增殖、凋亡的影响及其机制探讨

Effects of dichloroacetate on proliferation and apoptosis of bladder cancer T24 cells

目的观察二氯乙酸(DCA)对膀胱癌细胞株T24增殖、凋亡的影响,并探讨其相关机制。方法培养T24细胞并随机分成4个组,其中实验1组、实验2组和实验3组分别加入含5、10、20 mmol/L DCA的培养基,对照组加入等量含0.1%DMSO的培养基。分别于给药24、48、72 h后采用MTT法检测细胞增殖能力。给药48 h后,行Hoechst33258荧光染色观察并计数凋亡细胞,采用JC-1染色法检测细胞线粒体膜电位,采用Western blotting法检测各组细胞中的Bax、Bcl-2、Caspase-3蛋白。结果实验1、2、3组给药24、48、72 h细胞增殖能力均低于对照组,且随着DCA作用时间浓度增大、给药时间延长,细胞增殖抑制越明显(P均<0.05)。实验1、2、3组凋亡细胞数均高于对照组,线粒体膜电位均低于对照组(P均<0.05)。实验1、2、3组细胞中Bax、Caspase-3相对表达量高于对照组,Bcl-2相对表达量低于对照组(P均<0.05)。结论 DCA可抑制膀胱癌细胞株T24的增殖并诱导其凋亡,推测作用机制与线粒体膜电位降低、Bcl-2表达下调及Bax、Caspase-3表达上调有关。

Objective To observe the effects of dichloroacetate（ DCA） on the proliferation and apoptosis of bladder cancer cells T24 in vitro and to study the underlying mechanism. Methods The bladder cancer cells T24 were randomly divided into four groups： experimental group 1,experimental group 2,experimental group 3,and the control group. Cells in the experimental groups 1,2,and 3 were added with 5,10,and 20 mmol/L DCA,respectively; cells in the control group were added with 0. 1% dimethyl sulfoxide（ DMSO）. The cell proliferation of T24 cells were detected by MTT assay at 24,48,and 72 h after DCA treatment. At 48 h,the apoptosis was examined by Hoechst33258 staining,the mitochondrial membrane potential was detected by using JC-1 staining,and the expression levels of Bax protein,Bcl-2 protein and Caspase-3 protein were determined by Western blotting. Results The cell proliferation in the experimental groups 1,2,and 3 was lower than that of the the control group at 24,48 and 72 h,and with the time and increasing concentrations of DCA,the inhibition of cell proliferation was more significant（ all P〈0. 05）. The number of apoptosis cells in the experimental groups 1,2 and 3 was higher than that of the control group,and the mitochondrial membrane potential was lower than that of the control group（ all P〈0. 05）. Compared with the control group,the expression of Bax protein and Caspase-3 protein significantly increased,while the expression of Bcl-2 protein significantly decreased in the experimental groups 1,2,and 3（ all P〈0. 05）. Conclusions DCA can inhibit the proliferation and induce the apoptosis of bladder cancer cell line T24,which suggests that the mechanism may be related to the decrease of mitochondrial membrane potential level,upregulation of Bax protein and Caspase-3 protein,and down-regulation of Bcl-2 protein.