核因子-κB在TNF-α诱导HL-60细胞凋亡中的作用  

The Role of NF-κB in TNF-α Induced Apoptosis in Human Leukemic Cell Line HL-60

作　　者：唐爱萍[1] 陈卫华[1] 彭光洁 王开颜[1] 胡群[1] 刘双又[1] 张柳清 郭艺杰 王令仪 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科,武汉430030

出　　处：《华中科技大学学报（医学版）》2003年第1期77-80,121,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目 ( No. 3 9970 778)

摘　　要：目的　了解白血病细胞系 HL- 60中 ,肿瘤坏死因子 - α( TNF- α)诱导的核因子 - κB( NF- κB)活化与细胞凋亡之间的关系。方法　采用脂质体基因转染技术 ,将突变的核因子抑制蛋白 ( IκBα)质粒 ( p CMV4- IκBαS32 / 36 A)转染 HL-60细胞 ,于 48h后测其瞬时表达效应。用间接免疫荧光和 Western blot技术检测转染组和非转染组 HL- 60细胞的 NF-κB活化状态 ,同时用流式细胞术和 MTT法分别检测 TNF- α对两组 HL- 60细胞的凋亡诱导及生长抑制效应。结果　TNF- α可诱导非转染组 HL- 60细胞的 NF- κB活化 ,不能诱导转染组细胞的 NF- κB活化 ,即突变型 IκBα可特异性抑制HL- 60细胞的 NF- κB活化。结论　用突变型 IκBα特异性抑制 HL- 60细胞的 NF- κB活化 ,能明显增加其对 TNF-Objective To investigate the role of NF-κB in TNF-α induced apoptosis in HL-60. Methods A mutant IκBα (pCMV4-IκBα s32/36A) was transfected into HL-60 cells by lipofectin transfection technique and its transient effect was examined 48 h after transfection. The activation of NF-κB was detected by immune fluorescence cytochemistry and western blot methods with anti-p65 antibody. The apoptosis and inhibiting rate initiated by TNF-α in both transfected and untransfected HL-60 cells were measured respectively by a FACScan side scatter analysis and MTT method. Results TNF-α could activate NF-κB in untransfected cells but not transfected HL-60 cells. The percentages of apoptosis in transfected cells were significantly higher than in untransfected ones (P<0.01). And so did the inhibiting rate (P<0.01). Conclusion The resistance of HL-60 towards apoptosis induced by TNF-α can be mediated by NF-κB activation. The inhibition of NF-κB activation by mutant IκBα can attenuate the resistance of HL-60 cells and increase its sensitivity to TNF-α.

关 键 词：核因子-KB 核因子抑制蛋白 肿瘤坏死因子-α 细胞系 HL-60 细胞凋亡 

分 类 号：R730.3[医药卫生—肿瘤]