小剂量阿司匹林协同干扰素-α诱导肝癌BEL-7402细胞凋亡的实验研究  

The role and mechanism of low-dose aspirin and IFN-α in inducing hepatocellular carcinoma apoptosis in BEL-7402 cells

作　　者：董兆如[1] 李涛[1] 曲思凤[1] 罗黎希[1] 张春[1] 王钢普[1] 陈泽婷[1] 李小玮[1] 智绪亭[1] 

机构地区：[1]山东大学齐鲁医院普外科,济南250012

出　　处：《中华肝胆外科杂志》2012年第4期292-295,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：基金项目：国家自然科学基金（基金编号：30972889）;山东大学自主创新基金（基金编号：2010TS030）资助

摘　　要：目的探讨小剂量阿司匹林协同干扰素-α（IFN-α）诱导肝癌BEL-7402细胞凋亡的作用及机制。方法体外培养的人肝癌BEL-7402细胞经IFN-α单独或联合小剂量阿司匹林处理后，采用MTT法测定细胞的增殖情况；流式细胞术分析不同组药物对BEI。-7402细胞凋亡的影响，Western blot检测BEL-7402细胞凋亡相关蛋白表达的变化。结果MTT法检测显示，与对照组相比，作用48h时IFN-α组和阿司匹林组的肝癌细胞增殖率分别为82．45％±1．71％和83．22％±2．26％，联合用药组为69．84％±1．18％，联合用药组细胞增殖抑制率明显低于单独用药组（P〈0．05）；流式细胞术检测结果显示，IFN-α组和阿司匹林组细胞凋亡率分别为14．78％±1．93％和14．00％±0．610，联合用药组为21．68％±1．28％，明显高于单独用药组（P〈0．05）。Western blot检测发现IFN-α及小剂量阿司匹林可促进caspase-3及caspase-9的表达，而二者联合应用时caspase-3及caspase-9亦被明显激活。进一步检测显示，IFN-α单独或联合阿司匹林可促进肝癌细胞促凋亡蛋白Bax的表达（P〈0．05），而抗凋亡蛋白Bcl-2和Bcl—xl表达未见明显变化（P〉0．05）。结论小剂量阿司匹林可协同IFN-α抑制BEL-7402细胞的增殖，通过激活caspase-3及caspase-9诱导肿瘤细胞凋亡，该作用可能与促凋亡蛋白Bax表达的增高有关。Objective To investigate the role and mechanism ot low-dose aspmn concurrent with IFN-α in inducing hepatocellular carcinoma apoptosis in BEL-7402 cells. Methods BEL-7402 cells were cultured and treated with IFN-α, or low dose aspirin or both. MTT and flow cytometry were used to measure the cell proliferation and apoptosis after treatment with a singular drug or the combined regiment. The expressions of the apoptosis-related proteins were detected by Western blot. Results MTT assay revealed after IFN-α administration alone or combined with aspirin treatment for 48 h, the proliferation ratio of the IFN-α or aspirin group were 82.45% ± 1.71% and 83.22± 2.26 %, compared with the control group. The group which received the combined therapy had a proliferation ratio of 69.84% ±1.18%, which was significantly lower than the single groups （P〈0.05）. The flow cytometry revealed that the apoptosis ratio in IFN-α group and aspirin group were 14.78%± 1.93% and 14.00%±0.61%, respectively, while the IFN-α ＋ aspirin group was 21.68%±1.28%, which was also significantly higher than that of the single groups （P〈0.05）. Western blot detected that IFN-α and aspirin （1 mmol/L） could promote caspase-3 and caspase-9 protein expressions, and when the two drugs were combined, caspase-3 and caspase-9 were also significantly activated. IFN-α alone or combined with aspirin can promote the expression of pro-apoptotic protein Bax （P〈0.05）, while the anti-apoptotic proteins expression of Bcl-2 and Bcl-xl did not change significantly （P〈0.05）. Conclusions Low dose aspirin can cooperate with IFN-α in inhibiting the BEL-7402 cell growth and inducing the cell apoptosis by activating and increasing caspase 3 and caspase-9 levels, which may be related to the increased expression of pro-apoptotic protein Bax.

关 键 词：肝细胞癌 阿司匹林 干扰素-Α 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]