p33^(ING1b)基因对SW480细胞生长增殖的影响  

Effect on proliferation of SW480 cell line with p33^(ING1b) genetransfection

作　　者：赵帅[1,2] 贺修胜[1] 吕文玲 罗桥[1] 

机构地区：[1]南华大学肿瘤研究所 [2]长沙市第三医院病理科,湖南长沙410015 [3]长沙市第三医院病理科

出　　处：《中国癌症杂志》2008年第8期578-582,共5页China Oncology

基　　金：湖南省卫生厅Y02-069;省自然科学基金(NO:04JJ3059);湖南省教育厅课题(No:01B016)

摘　　要：背景与目的:p33ING1b基因作为一个新的候选抑瘤基因,具有多种生物学功能。本实验旨在研究p33ING1b基因对人结肠癌细胞SW480生长的影响。方法:经Western印迹和免疫细胞化学鉴定,通过生长曲线绘制、软琼脂集落形成实验和流式细胞仪分析检测p33ING1b基因转入对SW480细胞生长增殖以及细胞周期改变和凋亡率方面的影响,并用Western印迹法,检测3组细胞p53、p21WAF1、Bax及Bcl-2蛋白的表达情况,探讨p33ING1b基因抑瘤的可能分子机制。结果:与未转染组(SW480)和空载体组(pcDNA3.1(+)/SW480)相比,p33ING1b蛋白转染组(pcDNA3.1(+)/p33ING1b/SW480)细胞生长增殖速度减慢(P<0.05);软琼脂集落形成率降低(P<0.01);凋亡率增高(P<0.05),而对细胞周期的改变不明显(P>0.05)。p33ING1b基因在SW480细胞中高表达,能有效抑制SW480细胞的生长,并促进其凋亡。Western印迹法分析显示SW480细胞中p33ING1b基因高表达,导致Bax蛋白表达水平升高、Bcl-2蛋白质表达水平降低,差异有显著性(P<0.05);p53及p21WAF1蛋白表达水平稍有升高,但差异无显著性(P>0.05)。结论:高表达外源性p33ING1b基因后,SW480细胞生长增殖速度减慢,凋亡增加,其机制可能与Bax蛋白表达上调、Bcl-2蛋白表达下调有关。Background and purpose： As a new candidate tumor suppressor gene, p33^ING1b has many biological functions. This study was done to investigate its role in the regulation of the proliferation of human colon cancer cell line SW480. Methods： The pcDNA3.1（＋）/p33^ING1b/sw480 cells were identified by Western blot and S-P immunohistochemical method. In order to elucidate the effect of expression of exogenous p33^ING1b gene on the colorectal cancer cell SW480, the proliferation rates were analyzed by growth curves and colony formation assay in soft agar for cells including SW480, pcDNA3.1（＋）/p33^ING1b/sw480 and pcDNA3.1 （＋）/SW480. At same time, the apoptotic rate of cells and the cell cycle analysis were also tested by flow cytometry.Furthermore,using western blot analysis,we detected the expression level of the protein p53, p21^WAF1,Bax and Bcl-2 in those three group cells, which initially indicate the molecule mechanism of inducing apoptosis by gene p33^ING1b. Results： The cell growth rates of SW480 cells transfected with pcDNA3.1 （＋）/p33^ING1b were slower than those transfected with pcDNA3.1 （＋） or untransfected.The colony formation efficiency of pcDNA3.1（＋）/p33^ING1b/SW480 were decreased and the apoptotic rates were increased compared with pcDNA3.1（＋）/SW480 and SW480（P〈0.05）, which indicated that overexpression of p33^ING1b gene could efficiently inhibit the growth of SW480 cells and induce apoptosis, but the difference of cell cycle was not obvious. Ectopic overexpression of p33^ING1b up-regulated the expression of Bax remarkably and down-regulated the expression of Bcl-2 in SW480 cells （P〈0.05） detected by western blotting, but it had no significant impact on the expression of p53 and p21^WAF1 （P〉0.05）. Conclusion： After overexpression of exogenous p33^ING1b protein in SW480 cell, there were inhibition of the proliferation rate and induction of apoptosis, the molecular mechanism might be associated with upregulated expression of Bax and down-regulat

关 键 词：结肠癌SW480细胞系 p33^ING1b基因 转染 基因表达 

分 类 号：R730.231[医药卫生—肿瘤]