摘要
目的探讨过表达NOTCH-1对TNFα诱导的胃癌BGC-823细胞凋亡的调节作用。方法用包含NOTCH-1胞内段的反转录病毒载体转染BGC-823,G-418筛选后获得细胞克隆。TNFα作用于各组细胞后用MTT法、流式细胞术、W estern b lot及EMSA等检测细胞生长、凋亡、NOTCH-1表达、NF-κB及caspase-3活性。结果转染ICN的细胞中NOTCH-1及其靶基因HES-1表达强度明显强于对照组。经TNFα作用后,过表达NOTCH-1的胃癌细胞的细胞杀伤率、凋亡率显著低于对照组;过表达NOTCH-1能明显抑制TNFα诱导的caspase-3活化;然而,未发现过表达NOTCH-1对TNFα诱导的NF-κB活化有明显的影响。结论过表达NOTCH-1抑制TNFα诱导的BGC-823细胞增殖抑制与凋亡,可能通过非NF-κB依赖途径抑制caspase-3活化所致。
Objective sis of gastric carcinoma bined with intracellular To explore the regulatory function of over-expression of NOTCH-1 carcinoma domain of BGC-823 cells. Methods BGC-823 cells were infected NOTCH-1. The cell clones, into which objective genes TNFct-induced apopto- with retrovirus reeomhad been transfected,were obtained after screening with G-418. Before and after incubation with TNFα, MTT assay, flow cytometry, Western blot and EMSA were used to detect cell growth, apoptosis, expressions of NOTCH-1, NF-κB and caspase-3 activity. Results There was stronger expression of NOTCH-I and its target gene HES-1 in the cells transfeeted with NOTCH-1 gene than that of control. After TNFcx treatment, the rates of killing and apoptosis in NOTCH-1-transfected ceils were all lower than those in control cells. Moreover, we showed over-expression of NOTCH-1 suppressed caspase-3 activation induced by TNFct. However, TNFα-induced activation of NF-KB was not affected by over-expression of NOTCH-1. Conclusion Over-expression of NOTCH-1 significantly protects BGC-823 cells from apoptosis and growth suppression induced by TNFot. This effect is mediated, at least in part, through inhibition of easpase-3 activation independent of NF-κB.
出处
《基础医学与临床》
CSCD
北大核心
2009年第7期726-730,共5页
Basic and Clinical Medicine
基金
浙江省医药卫生科学研究基金(2008B204)
