增强p53、p21及抑制c-myc表达对MCF-7细胞增殖的协同抑制作用  

COORDINATION INHIBITION OF THE PROLIFERATION OF MCF-7 BY ENHANCING EXPRESSION OF p53,p21 AND DECREASING EXPRESSION OF c-myc

作　　者：张海江[1] 王楠[1] 朱晓宇[1] 桑建利[1] 何大澄[1] 

机构地区：[1]北京师范大学生命科学学院细胞生物学研究所,北京100875

出　　处：《实验生物学报》2005年第1期16-22,共7页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(批准号:30270663)国家重点基础研究发展规划资助项目(批准号:G1999053901)

摘　　要：为了探讨增强p53、p21基因表达水平和降低c-myc基因表达水平对乳腺癌细胞MCF-7增殖的协同抑制作用,以及这些基因对细胞产生效应时的相互关系,本研究中首先构建了正义的p53、p21和反义的c-myc3种真核细胞表达载体,并根据析因实验设计三种载体不同剂量组合。按照组合用质粒转染细胞,然后对转染细胞的增殖抑制率进行检测,并采用金正均Q值法、单因素方差分析中的LSD法、聚类分析法等统计学方法对结果进行统计分析。结果显示,不同量的p53、p21反义c-myc对MCF-7细胞的增殖均有抑制作用,抑制的程度各基因间存在差异。在各基因组合中,p21与反义c-myc,p53与反义c-myc联用具有协同作用,对MCF-7细胞的增殖产生更强的抑制,而p53与p21之间未显示出协同作用。对三基因协同结果进行聚类分析后,发现第一类组合协同作用最明显,第九类组合的抑制率最高。由此推测,作为抑癌基因的p53或CDK抑制基因p21高表达,同时原癌基因c-myc表达受到抑制,可相互协同显著增强对MCF-7细胞增殖的抑制作用。To study the coordination inhibition of the proliferation of breast cancer cell line MCF-7 by enhancing expression of p53,p21 and decreasing expression of c-myc, as well as the relationship among these genes when they express in the cells, eukaryotic expression plasmids of sense p53,p21 and antisense c-myc were constructed first, different concentrations of three plas- mids were determined and the combinations were designed according to factorial design. Cells were transfected by the combination plasimids, then the inhibition rate of proliferation of the transfected cells were tested. The results were analyzed using the methods of statistics including Jin's Q-test, Least significant difference (LSD),cluster analysis. It was shown that all of them (Sense p21,p53,antisense c-myc) could inhibit the proliferation of MCF-7 in different levels of concentration, while the degrees of inhibition were different among them. In the aspect of coordi- nation inhibition, the proliferation of MCF-7 was inhibited much strongly by p21 coordination with antisense c-myc and p53 coordination with antisense c-myc, but p53 coordination with p21 did not show inhibition effect. Rusults of cluster analysis showed that the first combination of three genes showed the best coordination, the ninth combination had the highest rate of inhibition. In conclusion, we suppose that when p53 as a cancer suppressor gene and p21 as a CDK sup- pressor gene express highly, and the expression of c-myc as a cancer gene is inhibited meantime could obviously enhance the inhibition on the proliferation of MCF-7 cells by coordination effect.

关 键 词：c—myc MCF-7细胞 增殖 反义 抑制作用 真核细胞表达载体 基因 转染细胞 表达水平 质粒 

分 类 号：R73-3[医药卫生—肿瘤]