论文部分内容阅读
目的 观察ｃｙｃｌｉｎ Ｅ 的高表达对乳腺癌细胞ＭＣＦ７ 生长及周期的影响。方法 构建ｃｙｃｌｉｎ ＥｃＤＮＡ真核表达载体并采用ｌｉｐｏｆｅｃｔＡＭＩＮＥ 转染方法将其导入ＭＣＦ７ 细胞，获得稳定表达ｃｙｃｌｉｎ Ｅ的细胞系。通过对细胞生长曲线绘制、３ＨＴｄＲ测定及细胞周期分布等的分析，观察其对细胞生长、增殖的影响。结果 ｃｙｃｌｉｎ Ｅ的高表达可以使细胞的生长速度加快（ 约为对照１－５２ 倍） 及ＤＮＡ 掺入增加（４－２ 倍） ，Ｇ１Ｓ移行加速；ｐＲＢ的磷酸化形式的增多（３ 倍） 。结论 ｃｙｃｌｉｎ Ｅ的高表达可明显影响ＭＣＦ７ 细胞的生长、增殖，并可能通过ｐＲＢ 磷酸化，影响细胞周期Ｇ１Ｓ期移行而实现其对生长的调节。
Objective To observe the effect of high expression of cyclin E on the growth and cycle of breast cancer cell MCF-7. Methods The eukaryotic expression vector of cyclin EcDNA was constructed and introduced into MCF-7 cells by lipofectAMINE transfection. The cell line with stable expression of cyclin E was obtained. Through the analysis of cell growth curve, 3H-TdR assay and cell cycle distribution, the effects on cell growth and proliferation were observed. As a result, high expression of cyclin E can accelerate the growth of cells (about 1-52 times the control) and DNA incorporation (4-2 times), accelerate the transition of G1-S, and increase the phosphorylation of pRB (3 times). ). Conclusion The high expression of cyclin E can significantly affect the growth and proliferation of MCF-7 cells, and it may affect the growth of MCF-7 cells through the phosphorylation of pRB, affecting the G1-S transition of cell cycle.