小檗碱对人肾细胞癌细胞增殖、凋亡、DNA断裂及损伤修复的影响

doi:10.6040/j.issn.1671-7554.0.2017.1202

摘要/Abstract

摘要： 目的探讨小檗碱(BBR)对人肾细胞癌(RCC)细胞生长及DNA断裂的影响。方法采用CCK-8法测定BBR在0~240 μmol/L不同浓度下对RCC细胞(A498、786-O)增殖的影响;根据CCK-8检测结果,将RCC细胞分为对照组(0 μmol/L)、30 μmol/L组和60 μmol/L组,应用AnnexinV-FITC/PI双染色法及流式细胞术检测BBR对RCC细胞凋亡的影响;Western blotting检测RCC细胞中凋亡执行因子前体和降解体(pro-caspase3、cleaved-caspase3)、组蛋白H2A变构体(γH2A.X)和KU70蛋白的表达。结果用BBR处理RCC细胞后,细胞增殖受到抑制,并呈时间及药物浓度依赖性,差异有统计学意义(A498: P<0.001; 786-O: P=0.002);有效浓度的BBR可增加RCC细胞凋亡率;30、60 μmol/L组较0 μmol/L组cleaved-caspase3均表达增加(A498: P=0.018、P<0.001; 786-O: P=0.038、P<0.001),γH2A.X均表达增加(A498: P<0.001、P<0.001; 786-O: P<0.001、P<0.001),而KU70均表达下降(A498: P=0.002、P<0.001; 786-O: P<0.001、P<0.001);60较30 μmol/L组cleaved-caspase3表达增加(A498: P=0.020; 786-O: P=0.010),γH2A.X表达增加(A498: P=0.002; 786-O: P<0.001),而KU70下调(A498: P<0.001; 786-O: P=0.005)。结论 BBR可抑制人RCC细胞的增殖,诱导RCC细胞凋亡,并促使DNA断裂、抑制DNA的损伤修复。

关键词: 肾细胞癌, 小檗碱, 细胞凋亡, 细胞增殖, DNA损伤修复

Abstract: Objective To determine the effects of berberine(BBR)on the growth and DNA damage of human renal cell carcinoma(RCC)cells. Methods The effects of BBR on the proliferation of A498 and 786-O RCC cells at concentrations of 0-240 μmol/L were tested with CCK-8 testing kit. Based on the results of CCK-8 test, RCC cells were divided into control group(0 μmol/L), 30 μmol/L group and 60 μmol/L group. The cell apoptosis after BBR treatment was evaluated with Annexin V-FITC/PI Apoptosis Detection Kit. The expressions of proteins involved in the apoptotic pathway and DNA damage repair pathways, including pro-caspased3, cleaved-caspased3, γH2A.X and KU70, were assessed with Western blotting. Results The proliferation of RCC cells was significantly suppressed by BBR in a time- and dose-dependent manner(A498: P<0.001; 786-O: P=0.002). Cell apoptosis was obviously increased after BBR 山东大学学报 (医学版)56卷3期 -李孝峰,等. 小檗碱对人肾细胞癌细胞增殖、凋亡、DNA断裂及损伤修复的影响 \=-treatment at an optimal concentration and time point. In comparison with the control group, the 30 and 60 μmol/L groups showed increased expressions of cleaved-caspase3(A498: P=0.018, P<0.001; 786-O: P=0.038, P<0.001), and γH2A.X(A498: P<0.001, P<0.001; 786-O: P<0.001, P<0.001), but decreased expression of KU70(A498: P=0.002, P<0.001; 786-O: P<0.001, P<0.001). Compared with the 30 μmol/L group, the 60 μmol/L group showed increased expressions of cleaved-caspase3(A498: P=0.020; 786-O: P=0.010), and γH2A.X(A498: P=0.002; 786-O: P<0.001), but decreased expression of KU70(A498: P<0.001; 786-O: P=0.005). Conclusion BBR can effectively suppress RCC cell proliferation, induce RCC cell apoptosis and DNA damage, and inhibit DNA damage and repair.

Key words: Renal cell carcinoma, Berberine, Cell apoptosis, Cell proliferation, DNA damage and repair

中图分类号:

R737.11

李孝峰,杜晓益,刘海南,刘承,范医东. 小檗碱对人肾细胞癌细胞增殖、凋亡、DNA断裂及损伤修复的影响[J]. 山东大学学报 (医学版), 2018, 56(3): 54-59.

LI Xiaofeng, DU Xiaoyi, LIU Hainan, LIU Cheng, FAN Yidong. Berberines effects on the proliferation, apoptosis and DNA damage and repair of human renal cell carcinoma cells[J]. Journal of Shandong University (Health Sciences), 2018, 56(3): 54-59.

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