蟾毒灵对CAPAN-2胰腺癌细胞增殖和细胞周期的影响

目的 研究蟾毒灵在胰腺癌CAPAN-2细胞系中的抗肿瘤作用，探讨其在胰腺癌中的抗肿瘤分子机制。方法 CCK-8法测定蟾毒灵对人胰腺癌CAPAN-2细胞增殖的影响；流式细胞术(FCM)检测蟾毒灵对细胞凋亡和细胞周期的影响；Western blotting法检测蟾毒灵作用于CAPAN-2细胞后Caspase-3酶原(pro-caspase-3)、Bcl-2、Bax、CDC25C、cyclinB1、CDC2蛋白表达水平的变化情况。结果 蟾毒灵对胰腺癌CAPAN-2细胞有抑制增殖的作用，并呈现出时间和浓度依赖性；蟾毒灵未能诱导CAPAN-2细胞凋亡，但经蟾毒灵处理后，CAPAN-2细胞周期被阻滞在G2/M期；经蟾毒灵处理后，CAPAN-2细胞中CDC25C、cyclinB1、CDC2等关键周期蛋白的表达量显著下降，但是pro-caspase-3、Bcl-2、Bax等凋亡相关蛋白的表达没有发生明显改变。结论 蟾毒灵可显著抑制胰腺癌CAPAN-2细胞增殖，通过诱导G2/M细胞周期阻滞而非诱导细胞凋亡，在CAPAN-2细胞中发挥抗肿瘤作用。

Effects of bufalin on the proliferation and cell cycle of human pancreatic cancer CAPAN-2 cells

Objective To investigate the anti-cancer role of bufalin in human pancreatic cancer CAPAN-2 cells and explore the underlying mechanisms. Methods The proliferation-inhibiting effect of bufalin on CAPAN-2 cells was measured by CCK-8 assay; the cell cycle distribution and apoptosis were detected by flow cytometry (FCM); the protein expression level of pro-caspase-3, Bcl-2, Bax, CDC25C, cyclinB1 and CDC2 were determined by Western blotting analysis. Results Bufalin inhibited the proliferation of CAPAN-2 cells in a time- and dose-dependent manner; bufalin induced G2/M phase arrest in CAPAN-2 cells but failed to induce cell apoptosis; the protein levels of CDC25C, cyclinB1 and CDC2 were dramatically down-regulated while there were no significant changes in pro-caspase-3, Bcl-2 and Bax protein levels. Conclusion Bufalin could inhibit the proliferation of CAPAN-2 cells by inducing G2/M phase arrest instead of apoptosis.