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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。
Objective To study the anti-tumor effect of bufalin in pancreatic cancer cell line CAPAN-2 and its anti-tumor molecular mechanism in pancreatic cancer. Methods CCK-8 method was used to determine the effect of bufalin on the proliferation of human pancreatic cancer cell line CAPAN-2. Flow cytometry (FCM) was used to detect the effect of bufalin on cell apoptosis and cell cycle. Caspase-3, Bcl-2, Bax, CDC25C, cyclinB1, CDC2 protein expression in CAPAN-2 cells. Results Bufalin inhibited the proliferation of pancreatic cancer cell line CAPAN-2 in a time-and concentration-dependent manner. Bufalin did not induce CAPAN-2 cell apoptosis. However, after treatment with bufalin, CAPAN-2 The cell cycle was arrested in G2 / M phase. The expression of CDC25C, cyclinB1, CDC2 and other key proteins in CAPAN-2 cells decreased significantly after treated with bufalin, but the expression of pro-caspase-3, Bcl- Apoptosis-related protein expression did not change significantly. Conclusions Bufalin can significantly inhibit the proliferation of pancreatic cancer cell line CAPAN-2 and induce apoptosis in G2 / M cell cycle but not in CAPAN-2 cells.