Genistein 对K562细胞生长及CyclinD1基因和bcr-abl融合基因表达的影响

目的 研究Genistein抑制K562细胞生长的机制及对CyclinD1基因和bcr-abl融合基因表达的影响。方法 用RT－PCR方法检测Genistein作用于K562细胞前后CyclinD1基因和bcr-abl融合基因的mRNA表达变化，用免疫荧光、流式细胞仪和电镜检测CDK4蛋白的表达、细胞周期及凋亡。结果 K562细胞中CyclinD1基因、bcr-abl融合基因的mRNA和CDK4蛋白表达阳性，用Genistein与K562细胞共同孵育后，bcr-abl融合基因的mRNA表达阴性，CyclinD1的mRNA和CDK4蛋白表达下降，细胞阻滞于G2／M期，K562细胞出现凋亡。结果 bcr-abl融合基因、CyclinD1基因和CDK4蛋白在K562细胞中高表达，Genistein能使K562细胞生长受抑，诱导其调亡，抑制bcr-abl基因的mRNA表达，并使CyclinD1基因的mRNA和CDK4蛋白表达下降，表明bcr-abl融合基因、CyclinD1基因和CDK4对慢粒发展存在内在关系。

Growth of K562 cells and the expression of CyclinD1 gene and bcr-abl fusion gene effected by Genistein

AIM To investigate the mechanism of how Genistein restrains the growth of K562 cells, and its effect on the expression of CyclinD1 gene and bcr abl fusion gene. METHODS Genistein was incubated with K562 cells, the mRNA of CyclinD1 gene and bcr abl fusion gene were examined by RT PCR and the expression of CDK4 protein, cell cycle and apoptosis were examined by immunofluorescence, flow cytometry and electron microscope at the same time. RESULTS The expression of CyclinD1 gene and bcr abl fusion gene' mRNA was positive in K562 cells. After K562 cells was incubated with Genistein, the expression of bcr abl fusion gene' mRNA and CDK4 protein were negative, mRNA of CyclinD1 gene decreased. And the cell was blocked in G2/M phase. Using electron microscope we could observe the cell's apoptosis. CONCLUSION CyclinD1gene and bcr abl fusion gene are highly expressed in K562 cells. The growth of K562 cells could be restrained by using Genistein, which induces K562 cell's apoptosis, repress the expression of bcr abl fusion gene, and makes CyclinD1 and CDK4 protein decrease. This illuminates that CyclinD1 gene, CDK4 protein and bcr abl fusion gene may have relation to the pathogenesis of chronic myelogenous leukemia (CML).