阿司匹林对人肾上腺皮质癌细胞增殖和凋亡作用的研究  

Effect of aspirin on the proliferation and apoptosis of human adrenal cortical cancer cells

作　　者：卢晓 邢增苗 卢德成[1] 梁杏欢[1] 黄振兴[1] 杨海燕[1] 罗佐杰[1] Lu Xiao, Xing Zengmiao , Lu Decheng , Liang Xinghuan, Huang Zhenxing , Yang Haiyan, Luo Zuojie(Department of Endocrinology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China)

机构地区：[1]广西医科大学第一附属医院内分泌科,南宁530021

出　　处：《中华内分泌代谢杂志》2018年第9期732-737,共6页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金（81060220）

摘　　要：目的 观察阿司匹林对人肾上腺皮质癌细胞增殖抑制作用及诱导细胞凋亡的影响,探讨其相关机制.方法 体外培养人肾上腺皮质癌SW-13细胞和H295R细胞,实验组为含不同浓度阿司匹林（终浓度为0.125、0.25、0.5、1 mg/ml）的DMEM/F-12完全培养基,对照组为不加入阿司匹林的含1%无水乙醇的DMEM/F-12完全培养基.经过不同浓度阿司匹林作用不同时间后（24、48、72 h）,采用噻唑蓝（ MTT）法检测阿司匹林对两株细胞的增殖抑制情况;用药48 h后使用倒置显微镜观察细胞的形态学变化;通过Hoechst33258荧光染色观察和计数凋亡细胞,应用流式细胞术检测细胞凋亡率的改变,Western印迹法检测Bcl-2和Bax基因的表达.结果 MTT结果提示,SW-13细胞和H295R细胞经不同浓度阿司匹林处理后,生长抑制率均比对照组高（P＜0.05）,且当作用时间相同、药物浓度增大时,抑制率增高（P＜0.05）,呈剂量依赖性;当药物浓度相同时,随着作用时间的延长,细胞生长抑制率也升高（P＜0.05）,呈时间依赖性.不同浓度阿司匹林处理细胞48 h后,细胞凋亡数、细胞凋亡率均高于对照组（P＜0.01）;Western印迹结果经灰度值分析显示,阿司匹林处理细胞48 h后,各实验组与对照组相比,Bax的表达增加,差异有统计学意义（ P＜0.05）;相反,Bcl-2的相对表达量低于对照组（P＜0.05）.结论 阿司匹林能抑制肾上腺皮质癌细胞增殖并诱导其凋亡,呈剂量和时间依赖性,可能的机制是上调Bax表达并下调Bcl-2表达.Objective To observe the effect of aspirin on the proliferation inhibition and apoptosis of human adrenal cortical cancer cells, and to explore preliminarily the related mechanisms. Methods The human adrenal cortical cancer cells were cultured in vitro. The experimental group was DEME/F-12 complete medium which contained different concentrations of aspirin （final concentration of 0.125, 0.25, 0.5, 1 mg/ml）. The control group was DEME/F-12 complete medium which had no aspirin but 1%anhydrous ethanol instead. After treatment by aspirin at different concentrations for different durations （24, 48, 72 hours）, we detected the proliferation inhibition of SW-13 cells and H295R cells by methyl thiazolyl tetrazolium （MTT） method, observed the changes of cell morphology with the inverted microscope; Observed and counted apoptotic cells through Hoechst33258 fluorescent staining, tested the cell apoptosis by flow cytometry, and detected the expression of Bcl-2 and Bax by western blotting. Results The date of MTT showed that after treated by different concentrations of aspirin,the growth inhibition ratio of SW-13 cells and H295R cells were higher than the control group （P＜0.05）, and at the same time, the inhibition ratio would increase when the drug concentration increased （ P＜0. 05 ）, with a dose-dependent tendency. When the drug concentration was constant, the inhibition ratio increased with the duration of the drug （P＜0.05）, which showed a time-dependent tendency. The number of apoptosis cells and the cell apoptosis rate of both SW-13 and H295R which were treated by different concentrations of aspirin for 48 hours were higher than the control group （ P＜0. 01）. According to the analysis of grayscale value of western blotting, Aspirin can increase the expression of Bax （ P＜0.05）. On the contrary, the expression of bcl-2 in the experimental group was lower than that in the control group （P＜0.05）. Conclusion Aspirin may inhibit the growth of human adrenal cortical cancer cell in v

关 键 词：阿司匹林 SW-13细胞 H295R细胞 增殖抑制 细胞凋亡 

分 类 号：R736.6[医药卫生—肿瘤]