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抑制cyclinD1表达对乳腺癌细胞增殖及cyclinE CDK2和p21cip1转录的影响 |
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桑建利 叶颖江 王永潮
【摘要】 目的 分析探讨人乳腺癌细胞中cyclinD1的表达受到反义RNA抑制后，细胞增殖能力的变化以及cyclinE,CDK2和p21cip1(cip1／waf1／sdi1)基因表达受到的影响。 方法 将表达cyclinD1反义RNA的重组质粒转入人乳腺癌细胞，由此抑制细胞中cyclinD1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 cyclinD1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6d时，生长抑制率为54%。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。cyclinE和CDK2的mRNA水平显示出有不同程度的下降，而p21cip1的表达无明显变化。 结论 cyclinD1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cyclinE和CDK2的表达与cyclinD1的表达密切相关，而p21cip1的表达不受cyclinD1的影响。
【关键词】 乳腺癌；细胞增殖；cyclinD1；CDK2；p21cip1
【中图分类号】 Q28；R329 【文献标识码】 A 【文章编号】 0529-1356(2000)03-221
INHIBITION EFFECT OF cyclinD1 ON CELL PROLIFERATION AND
TRANSCRIPTION OF cyclinE, CDK2 AND p21cip1
IN HUMAN BREAST CANCER CELL LINE
SANG Jian-li YE Ying-jiang WANG Yong-chao
Key Laboratory of Cell Proliferation and Regulation Biology, College of Life Science,Beijing Normal University, Beijing 100875, China
General Surgerg,the Second Hospital,Beijing Medical University,Beijing 100044, China
【Abstract】 Objective To study the changes of cell proliferation and the effects on transcription of cyclinE, CDK2 and p21cip1(cip／waf1／sdil) when cyclinD1 was inhibited by antisense RNA in human breast cancer(Bcap-37). Methods The recombinant vector expressing cyclinD1 antisense RNA was transfected into the cells to inhibite the expression of cyclinD1, then the rate of cell proliferation and distribution of different phase of cell cycles were measured. At the same time the transcription of cyclinE, CDK2 and p21cip1 were analyzed by Northern blot. Results As comparing the cells in which cyclin D1 was inhibited with control cells, the cell proliferation reduced markedly, the radio of growth inhibition was about 54% at the sixth day in assay of cell growth. The changes of distribution of phases in cell cycle were oberved. The proportion of cells in G1 phase increased while the proportion of cells in S and G2／M phase decr[1] [2] [3] 下一页
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