中药千金子对肾癌细胞株786-0增殖作用的影响及对G250蛋白影响研究  

Study on effect and mechanism of Chinese medicine Qianjinzi on the proliferation of renal carcinoma cell line 786-0 and G250 protein

作　　者：董建设[1] 赵俊峰[1] 陈瑞廷[1] 张林超[1] 孙继建 潘世杰[1] 杨锦建[2] 

机构地区：[1]河南省中医院泌尿外科,河南郑州450008 [2]郑州大学第一附属医院泌尿外科,河南郑州450052

出　　处：《临床研究》2017年第5期3-6,共4页Clinical Research

摘　　要：目的 探讨中药千金子对肾癌细胞株786-0增殖作用的影响及对G250蛋白的影响研究.方法 选用肾癌细胞株786-0,分为A组（对照）、B组（5 mg/mL）、C组（10 mg/mL）、D组（20 mg/mL）,各组分别采用不同浓度千金子水溶液处理786-0细胞株.MTT比色法检测细胞增殖抑制率,Hoechst33258染色法观察细胞凋亡形态,流式细胞术检测细胞凋亡及细胞周期变化,western blot法检测细胞Fas、Caspase3、Caspase7、G250蛋白表达水平,RT-PCR法检测细胞G250 mRNA表达水平.结果 ①各浓度千金子水溶液在24、48、72h对786-0细胞增殖抑制率较高,差异有统计学意义（P〈0.05）,且浓度越高、作用时间越长,对细胞增殖的抑制作用越强.②B、C、D组细胞核均有明显的凋亡形态学变化.③与A组比较,C、D组细胞凋亡率较高（P〈0.05）,C、D组G1期细胞比例较高（P〈0.05）,C、D组S期及G2期细胞比例较低,差异有统计学意义（P〈0.05）.④与A组比较,B、C、D组Caspase3、Caspase7蛋白表达水平较高（P〈0.05）,C、D组Fas蛋白表达水平较高,差异均有统计学意义（P〈0.05）.⑤与A组比较,B、C、D组G250 mRNA及G250蛋白表达水平较高,差异有统计学意义（P〈0.05）.结论 中药千金子对肾癌细胞株786-0增殖有明显的抑制作用,其机制为上调G250抗原及Fas、Caspase3、Caspase7蛋白表达,促进细胞发生G1期阻滞,引起786-0细胞凋亡.Objective To analysis the effect and mechanism of chinese medicine Qianjinzi on the proliferation of renal carcinoma cell line 786-0 and G250 protein. Methods Renal carcinoma cell line 786-0 Were selected, divided into A group （control）, B group （Smg/mL）, C group （10 mg/mL）, D group （20mg/mL）, 786-0 cell lines were treated with different concen- tration of Qianjinzi solution in each group. The cell proliferation inhibition rate was detected by MTF colorimetric assay, the apoptotic morphology was observed by Hoechst33258 staining method, flow cytometry was used to detect cell apoptosis and cell cycle change, the expression levels of Fas, Caspase3, CaspaseT, G250 were detected by western blot method, the expres- sion level of G250mRNA was detected by RT-PCR method. Results ① The inhibition rate of 786-0 cells after 24, 48, 72 h were higher in all concentrations of Qianjinzi water solution, the difference was statistically significant （P〈0.05）. ② The nu- cleus of B, C, D group had obvious morphological changes of apoptosis. ③ The apoptosis rate and the proportion of G1 phase cells of C and D group were significantly higher than those of the A group （P〈0.05）, the proportion of S phase and G2 phase cells were significantly lower in C and D group than those in A group （P〈0.05）. ④ The expression levels of Caspase3,Cas- pase7 protein were significantly higher in B, C, D group than those in A group （P〈0.05）, the expression level of Fas protein was significantly higher in C, D group than that of A group （P〈0.05）. ⑤ The expression levels of G250 mRNA and G250 pro- tein were significantly higher in B,C,D group than those in A group （P〈0.05）. Conclusion The chinese medicine Qianjinzi has a significant inhibitory effect on renal carcinoma cell line 786-0, the mechanism was to up regulate the expression of G250 antigen and Fas, Caspase3, Caspase7 protein, to promote the occurrence of G1 block and induce apoptosis of 786-0 cells.

关 键 词：千金子 肾癌细胞株786-0 细胞增殖 细胞周期 G250抗原 

分 类 号：R730.2[医药卫生—肿瘤]