双氢青蒿素对人胰腺癌细胞系增殖和凋亡的影响

目的探讨双氢青蒿素对人胰腺癌细胞(PANC-1)增殖活性及凋亡的影响,及其作用机制。方法30、60、120、240和480μmol/L双氢青蒿素作用PANC-1 48 h后,采用MTT法检测双氢青蒿素对PANC-1增殖活性的影响,流式细胞术检测其细胞周期和凋亡的变化,Western blotting检测细胞内周期相关蛋白cyclin B1、Cdk1、p21及凋亡相关蛋白Bcl-2,Bax,Caspase-3,Caspase-9和细胞色素C(Cyto C)蛋白表达变化。结果 MTT实验结果表明,30~480μmol/L双氢青蒿素对PANC-1增殖活性具有明显的抑制作用(P<0.05)。流式细胞仪检测结果显示,双氢青蒿素可以使PANC-1细胞周期阻滞于G2/M期,并诱发PANC-1细胞发生凋亡,且随药物浓度呈现升高趋势(P<0.05)。Western blotting结果显示,双氢青蒿素作用后周期相关蛋白Cdk1和Cyclin B1蛋白表达降低,而P21蛋白表达升高。凋亡相关蛋白Bcl-2蛋白表达下调,Bax蛋白表达上调,Bax/Bcl-2蛋白表达比例升高,cleaved Caspase-3,cleaved Caspase-9和Cyto C蛋白表达升高。结论双氢青蒿素能抑制PANC-1增殖并诱发细胞凋亡,其凋亡机制可能与线粒体途径相关。

Effects of dihydroartemisinin on proliferation and apoptosis of human pancreatic cancer cell line

Objective To investigate the effect of dihydroartemisinin on proliferation and apoptosis of human pancreatic cancer cell (PANC-1) and explore its mechanism. Methods After PANC-1cells treated with 30, 60, 120, 240 and 480 μmol/L dihydroartemisinin for 48 hours, MTT assay was used to detect the proliferation effect of dihydroartemisinin on PANC-1 cells. The cell cycle and apoptosis were detected by flow cytometry. The expressions of cyclin B1, Cdk1, p21 and apoptosis related protein Bcl-2, Bax, Caspase-3, Caspase-9 and cytochrome C(Cyto C) were detected by Western blotting. Results MTT result showed that the proliferation of PANC-1 cells was inhibited obviously after treatment with dihydroartemisinin by the concentration of 30-480 μmol/L (P<0.05). The result of flow cytometry showed that dihydroartemisinin could block PANC-1 cell cycle in G₂/M phase and induce cells apoptosis (P<0.05). The result of Western blotting showed that the expressions of Cdk1 and cyclin B1 were decreased and the expression of P21 increased after treatment with dihydroartemisinin. The expression of Bcl-2 was down regulated, expression of Bax was up-regulated, the ration of Bax/Bcl-2 was increased and the expression of cleaved Caspase-3, cleaved Caspase-9 and Cyto C were up-regulated. Conclusion Dihydroartemisinin can inhibit the proliferation of PANC-1 cells and induce apoptosis. The possible mechanism might be related to mitochondrial pathway.