活化的Notch1下调NF-κB对宫颈癌细胞增殖及凋亡的影响  

Impact of downregulation of NF-kB by activated Notch1 on proliferation and apoptosis of cervical carcinoma cells

作　　者：姚军[1] 钱翠娟[2] 伍欣星[3] 

机构地区：[1]台州学院医学院,浙江台州318000 [2]台州市立医院,浙江台州318000 [3]武汉大学医学院病毒研究所,湖北武汉437000

出　　处：《中国热带医学》2009年第4期632-635,共4页China Tropical Medicine

摘　　要：目的构建重组逆转录病毒表达载体pCLNRX-ICN,研究Notch1过度活化对人宫颈癌HeLa细胞生长的影响,并探讨其作用机制。方法通过逆转录病毒感染,将Notch1(ICN)基因导入并整合到HeLa细胞的基因组中。MTT法检测稳定表达ICN对HeLa细胞生长的影响,并将阳性克隆细胞皮下接种裸鼠,观察ICN基因转染前后细胞系在裸鼠体内的致瘤能力,流式细胞仪分析细胞凋亡的变化,Western blot及EMSA检测转录因子NF-κB及相关基因的变化。结果成功建立了稳定表达ICN基因的宫颈癌细胞株HeLa-ICN。活化的Notch1显著抑制HeLa细胞体内外增殖能力。而且在诱导HeLa细胞发生凋亡的同时,活化的Notch1可上调HeLa细胞中IκBα的表达,并下调NF-κB活性及其调节基因Bcl-2、c-Myc和cyclin D1的表达。结论活化的Notch1可通过诱导细胞凋亡和下调NF-κB活性而抑制人宫颈癌细胞增殖。Objective To construct the vector of pCLNRX-ICN for expression of recombinant retroviruses amd investigate the impact of overactivation of Notch1 on the growth of Hela cell in cervical cancer patients. Methods The recombinant retroviruses were used to infect HeLa cells. After infected, proliferation of HeLa cells was observed by MTT assay. The growth of HeLa cells in vivo were observed through the nude mice animal model.Apoptosis of HeLa cells were tested using Hoechst 33342 staining and flow cytometry. NF-κB activation and its related genes were detected by EMSA and Western blot. Results HeLa-ICN cells, Notchl overexpressing cells, were successfully set up. After infection of recombinant retrovirus, the growth of HeLa cells was inhibited significantly. Morever, activated Notchl induced apoptosis in HeLa cells. Also, up-regulation of IκBα and down-regulation of Bcl-2, c-Myc and cyclin D1, companied by decrease in NF-κB activation, were obseved in HeLa-ICN cells. Conclusion Activated Notchl could inhibit the growth of cervical carcinoma cells through induction of apoptosis while deduction of NF-κB signaling.

关 键 词：宫颈癌细胞 NOTCH1 重组逆转录病毒载体 NF-ΚB 细胞凋亡 

分 类 号：R711.71[医药卫生—妇产科学]