Nrf3基因对大肠癌LoVo细胞系生长的影响  

Effect of Nrf3 on colorectal cancer LoVo cell line in vitro

作　　者：王强[1] 李晓利[1] 白雪娟[1] 黄仲曦[2] 崔春平[3] 丁彦青[2] 张金萍[4] 姚开泰[2] 许红民[5] 

机构地区：[1]解放军总医院309临床部器官移植中心,北京100091 [2]南方医科大学病理解剖教研室及肿瘤研究所,广州510515 [3]军事医学科学院放射医学研究所,北京100850 [4]解放军第305医院,北京100017 [5]解放军总医院病理科,北京100853

出　　处：《军医进修学院学报》2008年第4期262-264,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：全军"十五"医学科研基金重点项目(04Z041);北京市自然科学基金资助项目(7052064)

摘　　要：目的:在体外检测Nrf3基因对大肠癌LoVo的生长影响作用。方法:构建其融合蛋白真核表达载体,脂质体介导该真核表达载体转染大肠癌LoVo细胞系,FCM观察其在体外对大肠癌LoVo细胞系细胞周期和凋亡的影响;荧光显微镜观察候选基因亚细胞定位。结果:构建融合蛋白真核表达载体pEGFP-N1-Nrf3。RT-PCR方法检测Nrf3的表达,与芯片检测结果基本一致。荧光显微镜下观察Nrf3定位于细胞核内。FCM分析显示Nrf3影响下LoVo G2/M+S期细胞所占比例较对照组明显减少,G0/G1细胞所占比例明显增加。证实Nrf3可抑制大肠癌LoVo细胞的DNA合成和有丝分裂,促使细胞阻滞于G0/G1期,抑制大肠癌LoVo细胞的体外生长。FCM分析显示Nrf3在体外对大肠癌LoVo细胞的凋亡无影响。结论:Nrf3在体外具有抑制大肠癌增殖的功能,对大肠癌的细胞凋亡无影响,为肿瘤抑制基因。Objective： To observe the effect of Nrf3 on eoloreetal cancer LoVo cell line in vitro. Methods： The plasmid was constructed by the insertion of the candidate gene, and was transfeeted to LoVo cell line. The cell cycle and apoptosis of LoVo cell line were checked by flow eytometry（FCM） in vitro, and the subeellular localization of this candidate gene was observed by the fluorescence microscope. Results： The expression vector pEGFP-N1-Nrf3 was constructed. It expressed in 3 CRC and did not express in paired normal mueosa tissue. Nrf3 was expressed in LoVo cell line and located in cell nucleus. The ratio of LoVo cell in G2/M ＋ S was obviously reduced under the influence of Nrf3 by using FCM, and ratio of LoVo cell in GO/G1 was increased. DNA synthesis and mitosis of LoVo were restrained by Nrf3. The peak of deuto-G1 （ the peak of apoptosis） was not seen by FCM. So, we thought the apoptosis of LoVo cell line were not influenced by Ntis in vitro. Conclusion： As a tumor-suppress- ing gene, Nrf3 has the function of inhibiting cell proliferation of CRC in vitro, but not influence apoptosis.

关 键 词：肠肿瘤 Nrf3 质粒 流式细胞术 细胞周期 细胞凋亡 

分 类 号：R735.34[医药卫生—肿瘤]