木黄酮对宫颈永生化上皮细胞的影响及其对细胞周期的调控机制  

Effects of Genistein on immortalized cervical epithelial cells,and its regulation mechanisms in cell cycle

作　　者：陈庆云[1] 卞美璐[1] 刘军[1] 陈志华[2] 向青[2] 徐梅[2] 徐波[2] 

机构地区：[1]中日友好医院妇产科,北京100029 [2]中日友好临床医学研究所肿瘤与分子生物学室,北京100029

出　　处：《中日友好医院学报》2009年第2期98-101,共4页Journal of China-Japan Friendship Hospital

摘　　要：目的:观察木黄酮对人宫颈永生化上皮细胞生长的抑制作用,并探讨其调控细胞周期的相关机制。方法:本研究采用人宫颈永生化上皮细胞系(H8细胞),应用磺酰罗丹明B(SRB)法,绘制细胞生长曲线,获得抑制50%细胞生长所需药物浓度(IC50)。应用流式细胞仪分析细胞周期。免疫荧光法检测细胞周期调控因子cyclin D1,cyclin B1,CDK4,cdc2和p21的蛋白表达,RT-PCR半定量法检测其mRNA表达。结果:不同浓度的木黄酮对H8细胞有生长抑制作用,并呈剂量依赖关系。木黄酮对H8细胞作用d5的IC50为55μM。木黄酮作用后使H8细胞阻滞于G2/M期。木黄酮作用3d后,开始显著下调H8细胞cdc2的mRNA和蛋白表达(P<0.05),其对H8细胞cyclin D1、cyclin B1、CDK4和p21的mRNA和蛋白表达均无影响(P>0.05)。结论:木黄酮可以抑制宫颈永生化上皮细胞的生长,其作用机制之一可能是通过下调其cdc2的表达。Objective：To observe the effect of Genistein on human immortalized cervical epithelial cells and to investigate its regulation mechanisms in cell cycle.Methods：Human immortalized cervical epithelial cell line （HS） was used in this study.The effect of Genistein on H8 cells was tested in mono-layer culture,the growth curves and the concentrations required for 50% growth inhibition （IC50）at 5th-day were determined by the sulforhodamine B assay.Cellular DNA content was measured and analyzed using flow cytometry for cell cycle analysis.The immunological fluorescence was measured by flow cytometry for detecting protein expressions of cyclin D1,eyclin B1,CDK4,cde2 and p21 respectively and the mRNA expressions of them were determined by RT-PCR semi-quantitatively.Results：Genistein inhibited the growth of H8 eclls at different concentrations in a dose-dependent mode.The IC50 of Genistein at 5th day for H8 cells was 55μM.H8 cells treated by Genistein were accumulated in G2/M phase.After 3 days Genistein treatment,Genistein down-regulated the protein and mRNA levels of cdc2 in H8 cells.The expressions of cyclin D1,cyelin B1,CDK4 and p21 maintained no change after Genistein treatment in H8 cells.Conclusion：Genistein can inhibit the growth of cervical immortalized epithelial cells.One of it＇s mechanisms may be the down-regulation of expression of cdc2 in immortalized cervical cells.Genistein may be used as a cell cycle regulatory agent for chemoprevention against cervical cancer.

关 键 词：木黄酮 宫颈肿瘤 永生化 细胞周期 

分 类 号：R737.33[医药卫生—肿瘤]