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目的观察半夏生物碱对786-0细胞株的凋亡诱导作用,并探讨其可能作用机制。方法将786-0细胞株与半夏生物碱(400、200、100μg/ml)共同培养48 h;细胞计数试剂盒(cell counting kit-8,CCK-8)法测定细胞活性;流式细胞法检测细胞的凋亡率;蛋白免疫印迹法及实时定量聚合酶联反应(polymerase chain reaction,PCR)法检测细胞内葡萄糖转运蛋白78的表达;免疫组织化学法检测细胞中Bad的蛋白表达。结果与阴性对照组比较,干预组细胞活性下降(P<0.01)并发生明显的凋亡现象,差异具有统计学意义(P<0.05);免疫蛋白印迹法及实时定量PCR法检测发现,半夏生物碱干预各组中葡萄糖转运蛋白78表达明显下降(P<0.05);免疫组织化学结果提示,半夏生物碱亦能上调Bad蛋白在细胞内的表达。结论半夏生物碱能通过抑制葡萄糖转运蛋白78的表达,并能上调Bad蛋白的表达,从而促进人肾细胞癌的凋亡。
Objective To observe the apoptosis induction effect of Pinellia ternata alkaloids on 786-0 cell line and to explore its possible mechanism. Methods 786-0 cells were incubated with Pinellia ternata (400, 200, 100μg / ml) for 48 hours. The cell viability was measured by cell counting kit-8 (CCK-8) The apoptosis rate of cells was detected by Western blotting and real-time quantitative polymerase chain reaction (PCR). The expression of Bad protein was detected by immunohistochemistry. Results Compared with the negative control group, the cell viability in the intervention group decreased significantly (P <0.01) and obvious apoptosis occurred. The difference was statistically significant (P <0.05). The immunoblotting and real-time quantitative PCR Alkaloids significantly decreased the expression of glucose transporter 78 in each group (P <0.05). Immunohistochemistry showed that the alkaloids of Pinellia tuber further up-regulated the expression of Bad protein in the cells. Conclusion Pinellia alkaloids can promote the apoptosis of human renal cell carcinoma by inhibiting the expression of glucose transporter 78, increasing the expression of Bad protein.