论文部分内容阅读
目的:研究冬凌草甲素(oridonin,Ori)对人肾癌细胞的生长抑制、诱导凋亡作用及其机制。方法:MTT法检测Ori对密度为1×104/mL的人肾癌A-704细胞的生长抑制作用;流式细胞仪Annexin V-FITC/PI双染色法检测Ori作用于人肾癌A-704细胞24 h的细胞凋亡率;实时监测聚合酶链反应(Real-time PCR)法检测32μmol.L-1Ori作用人肾癌A-704细胞24h后的Bcl-2,Bax及Caspase-3基因表达水平的变化。结果:Ori可显著抑制人肾癌A-704细胞的生长,并呈明显的时间-效应关系和浓度-效应关系,64μmol.L-1Ori作用人肾癌A-704细胞60 h后抑制率为73%。随着Ori作用浓度增加,凋亡细胞数和坏死细胞数均增加,浓度越高,坏死细胞率增加越多,64μmol.L-1Ori作用人肾癌A-704细胞24 h后,坏死细胞升至32.4%。随着32μmol.L-1Ori作用时间延长,人肾癌A-704细胞Bax,Caspase-3基因的表达逐渐增强,Bcl-2基因的表达逐渐减弱(P均<0.05)。结论:Ori通过上调Bax基因和降低Bcl-2基因表达诱导细胞凋亡,从而抑制人肾癌A-704细胞的生长。
Objective: To investigate the effect of oridonin (Ori) on human renal cell carcinoma cell growth, induce apoptosis and its mechanism. Methods: The inhibitory effects of Ori on the proliferation of human renal carcinoma A-704 cells at a density of 1 × 104 / mL were determined by MTT assay. Ori on human renal carcinoma A-704 cells was detected by flow cytometry with Annexin V-FITC / PI double staining Cell apoptosis rate at 24 h. Real-time monitoring of Bcl-2, Bax and Caspase-3 gene expression in human renal cell carcinoma A-704 cells treated with 32 μmol·L-1 Ori was detected by Real-time PCR Horizontal changes. Results: Ori significantly inhibited the growth of human renal cell carcinoma A-704 cells and showed a time-effect relationship and concentration-effect relationship. The inhibition rate of 64-μmol·L-1 Ori on human renal cell line A-704 cells after 60 h was 73 %. As the concentration of Ori increased, the number of apoptotic cells and the number of necrotic cells increased. The higher the concentration of necrotic cells, the more necrotic cells increased. After treated with 64μmol·L-1Ori for 24 hours, the number of necrotic cells increased 32.4%. With the prolongation of 32μmol.L-1Ori, the expression of Bax and Caspase-3 genes in human renal cell carcinoma A-704 cells gradually increased, while the expression of Bcl-2 gene decreased gradually (all P <0.05). Conclusion: Ori can induce apoptosis of human renal cell carcinoma A-704 cells by up-regulating Bax gene and decreasing Bcl-2 gene expression.