CAJ | 학술논문

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BEL-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48 h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％±0.61％,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.
목적 탐토소제량아사필림협동간우소-α(IFN-α)유도간암BEL-7402세포조망적작용급궤제.방법 체외배양적인간암BEL-7402세포경IFN-α단독혹연합소제량아사필림처리후,채용MTT법측정세포적증식정황；류식세포술분석불동조약물대BEL-7402세포조망적영향,Western blot검측BEL-7402세포조망상관단백표체적변화.결과 MTT법검측현시,여대조조상비,작용48 h시IFN-α조화아사필림조적간암세포증식솔분별위82.45％±1.71％화83.22％±2.26％,연합용약조위69.84％±1.18％,연합용약조세포증식억제솔명현저우단독용약조(P＜0.05)；류식세포술검측결과현시,IFN-α조화아사필림조세포조망솔분별위14.78％±1.93％화14.00％±0.61％,연합용약조위21.68％±1.28％,명현고우단독용약조(P＜0.05).Western blot검측발현IFN-α급소제량아사필림가촉진caspase-3급caspase-9적표체,이이자연합응용시caspase-3급caspase-9역피명현격활.진일보검측현시,IFN-α단독혹연합아사필림가촉진간암세포촉조망단백Bax적표체(P＜0.05),이항조망단백Bcl-2화Bcl-xl표체미견명현변화(P＞0.05).결론 소제량아사필림가협동IFN-α억제BEL-7402세포적증식,통과격활caspase-3급caspase-9유도종류세포조망,해작용가능여촉조망단백Bax표체적증고유관.
Objective To investigate the role and mechanism of low-dose aspirin concurrent with IFN-a 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.