p53对人乳腺癌细胞MCF-7生长及周期相关基因的影响  

EFFECTS OF p53 ON GROWTH AND CYCLE RELATIVE GENES OF MCF-7 BREAST CANCER CELLS

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

机构地区：[1]北京师范大学细胞所细胞增殖与调控教育部重点实验室,北京100875

出　　处：《解剖学报》2005年第5期486-490,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30270663)

摘　　要：目的探讨抑癌基因p53对人乳腺癌细胞MCF-7生长以及周期相关基因cyclinD1、cyclinE、CDK2和CDK4表达的影响。方法将正义的p53真核表达载体导入MCF-7细胞,获得稳定表达正义p53的细胞模型,并经PCR和Western blot鉴定。分析了细胞的各种生长特性包括生长曲线、血清依赖、单细胞克隆形成、软琼脂成集落等;利用TdR双阻断法同步化细胞,流式细胞术分析了细胞周期;Western blot检测了其他周期相关基因的表达。结果p53的高表达可以使细胞的生长速度减慢、血清依赖性增加、单细胞克隆成集落能力降低、非锚定依赖性生长能力减弱,S期的进程减缓,一定程度上抑制cyclinD1和CDK2的表达,cyclinE和CDK4的表达未见明显变化。结论抑癌基因p53的表达能够抑制人乳腺癌细胞MCF-7的生长,并可能通过阻抑cyclinD1和CDK2的表达而阻抑细胞周期S期进程,实现其调节功能。Objective We investigated the effect of p53,a inhibitor of oncogene,on the growth of MCF-7 cells and the expression of some cell cycle relative gene in MCF-7 cells. Methods A eukaryotic expression vector containing p53 gene was infected into MCF-7 cells,thus a cell line that could express p53 gene steadily was identified by PCR and Western blot assay. We tested the cell growth capacity, serum dependent growth ability, plate colony rate and soft agar colony growth ability, and the cell cycle was analyzed by flow cytometry after a twice TdR cell cycle arrest. Finally, we tested the expression of some cell cycle relative genes by Western blot. Results The higher expression of p53 could result in a slower growth of the cell line, increasing of the serum dependent growth, reducing of the plate colony rate, increasing of the anchorage dependent growth, and delaying of entering the S phase. The expression of p53 gene could inhibit the expression of cyclinD1 and CDK2, while had little effect on the expression of cyclinE and CDK4. Conclusion The expression of p53 gene could inhibit the growth of the MCF-7 cell, and its function of cell cycle regulation is probably played by the dalaying of the S phase entering because of the inhibition of cyclinD1 and cyclinE gene expression.

关 键 词：乳腺癌 细胞周期 抑癌基因 P53 

分 类 号：Q785[生物学—分子生物学]