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目的探讨冬凌草甲素对胰腺癌PANC-1细胞生长抑制以及促进凋亡的机制。方法采用MTT还原法检测冬凌草甲素对PANC-1细胞生长的抑制作用;碘化丙啶(PI)染色和Hoechst33342染色法观察细胞形态学的变化;PI染色流式细胞术检测细胞周期分布;膜联蛋白V(Annexin V)/PI双标流式细胞术测定凋亡细胞比率;分光光度法检测Caspase-3酶活性;Western blotting法测定Caspase-3酶原(pro-caspase-3)、Bax和Bcl-2蛋白的表达变化。结果冬凌草甲素对胰腺癌细胞PANC-1具有明显的生长抑制作用,并呈现明显的剂量依赖性;细胞形态学观察发现,冬凌草甲素可诱导PANC-1细胞凋亡;细胞周期被阻滞在G2/M期,细胞凋亡率随药物浓度的增加而增加;Caspase-3酶原被激活;Bax蛋白表达量增加,Bcl-2蛋白表达无明显变化。结论冬凌草甲素可通过G2/M细胞周期阻滞和诱导凋亡抑制胰腺癌细胞PANC-1的生长,其分子机制可能是通过改变Bax/Bcl-2的表达比率,激活Caspase-3,从而促进PANC-1细胞发生凋亡。
Objective To investigate the mechanism of oridonin on pancreatic cancer PANC-1 cell growth inhibition and apoptosis. Methods MTT assay was used to detect the inhibitory effect of oridonin on the growth of PANC-1 cells. PI staining and Hoechst33342 staining were used to observe the morphological changes of cells. PI staining was used to detect cell cycle distribution ; The ratio of apoptotic cells was determined by Annexin V / PI double-flow cytometry; The activity of Caspase-3 was detected by spectrophotometry; The expressions of pro-caspase-3, Bax and Bcl-2 protein expression changes. Results Oridonin could significantly inhibit the growth of pancreatic cancer cell PANC-1 in a dose-dependent manner. Morphological observation showed that oridonin could induce the apoptosis of PANC-1 cells. The cell cycle Was arrested in G2 / M phase. The apoptotic rate increased with the increase of drug concentration. The activity of Caspase-3 was activated. The expression of Bax protein increased but the expression of Bcl-2 protein did not change significantly. Conclusion Oridonin can inhibit the growth of pancreatic cancer cell line PANC-1 through G2 / M cell cycle arrest and apoptosis induction. The molecular mechanism may be that Caspase-3 is activated by changing the expression ratio of Bax / Bcl-2, Thereby promoting the apoptosis of PANC-1 cells.