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目的 分析探讨人乳腺癌细胞中 cyclin D1的表达受到反义 RNA抑制后 ,细胞增殖能力的变化以及cyclin E,CDK2和 p2 1cip1 (cip1/ waf1/ sdi1)基因表达受到的影响。 方法 将表达 cyclin D1反义 RNA的重组质粒转入人乳腺癌细胞 ,由此抑制细胞中 cyclin D1的表达 ,然后分析细胞生长速率和各时相细胞分布比例的变化 ,并通过Northern blot分析有关基因的表达水平。 结果 cyclin D1表达受到抑制的细胞与对照相比 ,细胞增殖速率明显下降 ,培养至第 6 d时 ,生长抑制率为 5 4%。细胞周期各时相的分布比例也有较大的变化 ,G1期细胞比例上升 ,而 S期及 G2 / M期细胞比例下降。cyclin E和 CDK2的 m RNA水平显示出有不同程度的下降 ,而 p2 1cip1的表达无明显变化。 结论 cyclin D1表达的抑制可明显减弱人乳腺癌细胞的异常增殖 ,同时表明同为细胞周期调控基因的 cyclin E和CDK2的表达与 cyclin D1的表达密切相关 ,而 p2 1cip1的表达不受 cyclin D1的影响
Objective To investigate the effect of antisense RNA suppression on the expression of cyclin D1 in human breast cancer cells, the changes of cell proliferation and the effects of cyclin E, CDK2 and p21cip1 (cip1/waf1/sdi1) gene expression. METHODS: Recombinant plasmid expressing cyclin D1 antisense RNA was transfected into human breast cancer cells, thereby inhibiting the expression of cyclin D1 in the cells. Then the growth rate of cells and the proportion of cell distribution at each phase were analyzed, and the genes were analyzed by Northern blot. The level of expression. Results Compared with the control, the cell proliferation rate of cyclin D1 was significantly decreased, and the growth inhibition rate was 5 4% at the 6th day. The distribution ratio of the cell cycle at each phase also changed greatly. The proportion of cells in the G1 phase increased, while the proportion of cells in the S phase and the G2 / M phase decreased. The m RNA levels of cyclin E and CDK2 showed a different degree of decline, while p2 1cip1 expression did not change significantly. Conclusion The inhibition of cyclin D1 expression can significantly attenuate the abnormal proliferation of human breast cancer cells. Meanwhile, the expression of cyclin E and CDK2, which are both cell cycle regulators, is closely related to the expression of cyclin D1. The expression of p21cip1 is not affected by cyclin D1. influences