稳定表达Cyclin E细胞系的建立及Cyclin E高表达对人乳腺癌MCF 7细胞生长及细胞周期的影响

稳定表达Cyclin E细胞系的建立及Cyclin E高表达对人乳腺癌MCF 7细胞生长及细胞周期的影响

Building of Cell Line with Expression Cyclin E and Effect of Its Expression on Cells Growth and Cell Cycle Changes of Breast Cancer MCF 7 cell

摘要

摘要: 目的观察cyclin E 的高表达对乳腺癌细胞MCF-7 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MCF-7 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、3H-TdR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）,G1-S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MCF-7 细胞的生长、增殖，并可能通过pRB 磷酸化，影响细胞周期G1-S期移行而实现其对生长的调节。

Abstract: Objective To investigate the effect of cyclin E expression on cells growth and cell cycle changes of human breast cancer MCF 7 cell line. Methods The recombination eukryotic expression plasmid of cyclin E was constructed, and transferred into the MCF 7 cell line with lipofectAMINE.Cell clones were selected with G418.Cells growth properties, 3H TdR and cell cycle analysis were done. Results The cyclin E overexpression could obviously increase the cells growth speed (1 52 fold higher than vector control) and DNA synthesis (1 75 fold) and also increase the cells progression from G1 to S phase.The hyperphosphorylation form of pRB was increased (3 fold). Conclusion High expression of cyclin E could effect the cell growth and cell proliferation probably through regulating the phosphorylation of pRB and G1 S progression.

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