中药复方脏毒清体外诱导人结肠癌SW480细胞凋亡的影响

目的:探讨中药复方脏毒清对人结肠癌SW480细胞的促凋亡作用及机制。方法:体外培养人结肠癌细胞株SW480,用不同质量浓度的脏毒清(0.05,0.10,0.15,0.20,0.25 g·m L-1)进行干预12,24,36,48 h后,采用MTT法检测细胞增殖抑制率。用不同质量浓度脏毒清(0.10,0.15,0.20 g·m L-1)作用SW480细胞24 h后,另设空白组,采用用荧光染色技术和流式细胞仪检测细胞凋亡率,分光光度法测定半胱天冬酶-3(Caspase-3),Caspase-9的酶活性,Western blot技术检测凋亡蛋白Bax,Bcl-2蛋白的表达情况。结果:MTT结果显示脏毒清对其有明显的增殖抑制作用,并呈现时间和剂量依赖性;与空白组比较,脏毒清0.10,0.15,0.20 g·m L-1组能明显诱导SW480细胞凋亡,Caspase-3及Caspase-9酶活性明显升高,促凋亡蛋白Bax表达明显升高,抗凋亡蛋白Bal-2表达明显降低,均具有统计学意义(P0.01)。结论:脏毒清具有促进人结肠癌SW480细胞凋亡的作用,并通过线粒体凋亡途径发挥作用。

Effect on Apoptosis of Human Colon Cancer SW480 Cells by Traditional Chinese Medicine Compound of Zangduqing

Objective: To investigate the inducing apoptosis effect and the underlying mechanism of Zangduqing (ZDQ) on human colon cancer SW480 cells. Method: SW480 cells were cultured with different concentrations of ZDQ (0.05, 0.10, 0.15, 0.20, 0.25 g·mL^-1) for 12, 24, 36, 48 h in vitro. The cellular cytotoxicity was measured by using the MTT method. Then, SW480 cells were cultured with ZDQ (0.10, 0.15, 0.20 g·mL^-1) for 24 h in vitro and a control group was assigned. The apoptosis was detected by the DAPI and annexin V-FITC/propidium iodide assay. The activities of Caspase-3, -9 were assayed by using Caspase apoptosis detection kit. Protein levels of Bax and Bcl-2 were determined by using Western blot. Result: MTT showed that ZDQ had obvious inhibiting effect on the proliferation of human colon cancer SW480 cells with a dose- and time-dependent manner. Compared with the control group, the ZDQ could significantly inhibit the proliferation of SW480 cells, enhance the activity of Caspase-9 and Caspase-3, up-regulate the level of Bax, down-regulate the level of Bcl-2 (P<0.01). Conclusion: ZDQ could induce the apoptosis of SW480 cells obviously, which may be mediated by the mitochondrial apoptosis pathway.