抑瘤饮对胆管癌细胞增殖和凋亡的影响及机制

Effect and mechanism of Yiliuyin(Anti-tumor decoction) on proliferation and apoptosis of cholangiocarcinoma cells

目的观察中药复方抑瘤饮对胆管癌细胞增殖和凋亡的影响,探讨其可能的作用机制。方法取胆管癌QBC939细胞,随机分为抑瘤饮低、中、高浓度组和对照组,抑瘤饮低、中、高浓度组分别加入6、8、12 mg/m L抑瘤饮,对照组加入PBS,培养24 h。采用Hoechst 33258荧光染色法观察细胞形态学改变; CCK-8法测算各组培养24、48 h的细胞增殖抑制率;流式细胞仪测算细胞凋亡率; Western blotting法检测凋亡相关蛋白PARP、Caspase-9、Caspase-12、Bax、Bcl-2表达。结果与对照组比较,抑瘤饮高浓度组细胞呈明显的形态学改变,细胞核染色质固缩,核内荧光分布不均匀,出现畸形核。抑瘤饮各浓度组细胞增殖抑制率均高于对照组,其中抑瘤饮高浓度组细胞增殖抑制率高于低、中浓度组,各组48 h时的细胞增殖抑制率高于24 h(P均<0. 05)。抑瘤饮各浓度组细胞凋亡率均高于对照组(P均<0. 05)。抑瘤饮各浓度组PARP、Caspase-9、Caspase-12、Bax蛋白表达均高于对照组,Bcl-2蛋白表达均低于对照组(P均<0. 05)。结论抑瘤饮能够抑制胆管癌细胞增殖并诱导其凋亡,其机制可能与Caspase的内源信号凋亡途径有关。

Objective To study the effect of Fufang Yiliuyin(Anti-tumor decoction)on the proliferation and apoptosis of cholangiocarcinoma cells and to explore its possible mechanism.Methods The cholangiocarcinoma QBC939 cells were randomly divided into the low-,medium-,and high-concentration groups and control group;and the cells in the low-,medium-,and high-concentration groups of Yiliuyin were respectively added with 6,8,and 12 mg/mL Yiliuyin,whereas the cells in the control group were added with PBS,culturing for 24 h.The morphological changes of cells were observed by Hoechst 33258 fluorescence staining.The cell proliferation inhibition rate of each group at 24 and 48 h was detected by CCK-8.The apoptosis rate was detected by flow cytometry.The expression of apoptosis-related protein PARP,Caspase-9,Caspase-12,Bax,and Bcl-2 were detected by Western blotting.Results Compared with the control group,the cells in the high-concentration group of Yiliuyin showed significant morphological changes,such as the chromatin condensation of the nucleus,uneven fluorescence distribution in the nucleus,and malformed nuclei.The inhibition rate of cell proliferation in each concentration group was higher than that in the control group.The inhibition rate of cell proliferation in the high-concentration group was higher than that in the low-and medium-concentration groups.The inhibition rate of cell proliferation in each group at 48 h was higher than that at 24 h(all P<0.05).The apoptosis rate of each concentration group of Yiliuyin was higher than that of the control group(P<0.05).The expressions of PARP,Caspase-9,Caspase-12,and Bax in the concentration groups of Yiliuyin were higher than those in the control group(all P<0.05),while the expression of Bcl-2 protein was lower than that in the control group(P<0.05).Conclusion Yiliuyin can induce the apoptosis and inhibit the proliferation of cholangiocarcinoma cells and its mechanism may be related to the endogenous signaling apoptotic pathway of Caspase.