渥曼青霉素对化疗药物诱导BGC823细胞凋亡的影响

Effects of Wortmannin on Drug-Induced Apoptosis of Human Gastric Cancer Cell BGC823

目的:探讨磷脂酰肌醇3-激酶(PI3K)抑制剂渥曼青霉素(wortmannin)对足叶乙甙和阿霉素诱导的人胃癌细胞BGC823凋亡的影响。方法:常规培养人胃癌细胞BGC823,将细胞分为6组:①空白对照组(不加任何药物);②渥曼青霉素组(终浓度为40 nmol/L);③足叶乙甙组(终浓度为20μmol/L);④渥曼青霉素(终浓度为40nmol/L)+足叶乙甙组(终浓度为20μmol/L);⑤阿霉素组(终浓度为0.3μmol/L);⑥渥曼青霉素(终浓度为40nmol/L)+阿霉素组(终浓度为0.3μmol/L)。分别以上述药物干预24 h后,提取各组细胞的蛋白,以NaI法提取各组细胞的DNA。采用DNA琼脂糖凝胶电泳分析各组的细胞凋亡现象,用Western blot检测各组细胞中Caspase-3蛋白的活化表达。结果:DNA琼脂糖凝胶电泳示空白对照组细胞见较弱的DNA梯状谱(DNA Ladder),其他5组细胞均出现明显的DNA梯状谱;阿霉素与足叶乙甙干预后,胃癌细胞BGC823的Caspase-3活性较空白对照组增强,加用渥曼青霉素处理后,Caspase-3活性进一步增强。结论:PI3K抑制剂渥曼青霉素可增强化学治疗药物足叶乙甙和阿霉素对胃癌细胞的凋亡诱导作用,提高其化疗疗效。为寻求新的高效胃癌化疗方案提供理论依据。

Objective： To explore effects of Wortmannin, the specific inhibitor of the signaling pathway PI3K on apoptosis of human gastric cancer cell BGC823 induced by Etoposide and adriamycin. Methods： Gastric cancer cell BGC823 were cultured routinely, then divided into six groups： ①control group（not add any drugs）;②wortmannin group （40 nmol/L）;③etoposide group （20 μmol/L）; ④wortmannin （40 nmol/L） ＋ etoposide group （20 μmol/L）; ⑤adriamycin group （0.3 μmol/L） ; ⑥wortmannin （40 nmol/L） ＋ adriamycin group （0.3 μmol/L）. After the cells were treated for 24 hours respectively, the protein and DNA of each group was extracted. Apoptosis of gastric cancer cells was detected by DNA agarose gel electrophoresis. Caspase-3 protein expression was determined by Western bolt. Results. DNA agarose gel electrophoresis showed that apoptotic ladder in all groups were obvious except in control group. Western blot indicated that when the cells were treated with etoposide and adriamycin, Caspase-3 protein expression was increased, especially after treated with Wortmannin. Conclusion： PI3K inhibitor wortmannin may increase effects on apoptosis of BGC823 induced by chemotherapeutic agent, etoposide and adriamycin, and enhance their therapeutic effect. This may provide a new theory for increasing the therapeutic effect in gastric cancer treatment.