金雀黄素对HEC-1B细胞增殖及荧光素酶报告基因表达影响的研究*  

Influences of genistein on proliferation of HEC-1B and expressions of luciferase reporter genes

作　　者：刘小丽[1] 薛晓鸥[1] 唐炳华[1] 牛建昭[1] 

机构地区：[1]北京中医药大学东直门医院妇科,北京100700

出　　处：《北京中医药大学学报》2011年第4期236-240,共5页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家"十一五"科技支撑计划资助项目(NO.2003BA11B07-02);首都医学科技发展基金资助项目(NO.F-2007-III-11)

摘　　要：目的探讨金雀黄素对子宫内膜癌雌、孕激素受体阴性细胞系(HEC-1B细胞)增殖的影响及其受体作用机制,为临床运用金雀黄素抗肿瘤治疗提供理论依据。方法体外培养子宫内膜癌HEC-1B细胞,金雀黄素作用浓度分别为10×10-6、20×10-6、40×10-6、80×10-6、160×10-6mol/L,作用时间分别为24、48、72、96 h。采用四甲基偶氮唑盐(MTT)法检测金雀黄素对子宫内膜癌细胞增值的影响;同时采用荧光素酶报告基因的方法,检测不同浓度金雀黄素对雌激素应答原件(ERE)调控的雌激素受体α(ER-α)和雌激素受体β(ER-β)报告基因表达的影响,并比较其对ERα-和ER-β作用的差异性。结果金雀黄素对HEC-1B细胞体外增殖有明显抑制作用,差异有统计学意义(P<0.01),其抑制作用随药物作用浓度增加和作用时间的延长而逐渐增强,呈剂量及时间依赖性。在金雀黄素浓度为20×10-6、40×10-6mol/L的作用下,报告基因的表达较空白对照组明显提高,差异有统计学意义(P<0.01),且呈浓度依赖性;金雀黄素通过ER-β介导的报告基因表达水平的升高程度强于ER-α,差异有统计学意义(P<0.01)。结论金雀黄素可抑制子宫内膜癌细胞的增殖,这种抑制作用可能通过调节子宫内膜癌细胞ER-α和ER-β的表达,调整ER-β/ERα-比例而实现。Objective To investigate the influence and its receptor mechanism of genistein on the proliferation of negative cell line of estrogen receptor and progesterone receptor of endometrial carcinoma,HEC-1B and provide the theoretical base for the application of genistein in clinic.Methods HEC-1B were cultured in vitro,and than divided into the blank group,genistein group 1（10×10-6 mol/L）,genistein group 2（20×10-6 mol/L）,genistein group 3（40×10-6 mol/L）,genistein group 4（80×10-6 mol/L） and genistein group 5（160×10-6 mol/L） with different action time（24 hours,48 hours,72 hours and 96 hours）.The influence of genistein on the proliferation of HEC-1B was detected by using MTT method.The influences of genistein in different doses on the expressions of estrogen receptor-α（ER-α） and estrogen receptor-β（ER-β） regulated by estrogen response element（ERE） by applying the method of luciferase reporter gene.The difference between genistein effects on ER-α and ER-β were compared.Results The proliferation of HEC-1B in vitro was inhibited significantly by genistein with statistical difference（P0.05）.The inhibitory effect of genistein increased along with the increase of its concentration（10×10-6 mol/L-160×10-6 mol/L） and action time（24-72 hours） showing a dose-time dependency.The expressions of reporter genes were much higher in the genistein group 2 and 3 than those in the blank group（P0.05） showing a dose dependency.The increase of the expression of reporter gene mediated by ER-β was more significant that that by ER-α（P0.01）.Conclusion Genistein can inhibit the proliferation of endometrial carcinoma through regulating the expressions of ER-α and ER-β,and proportion of ER-β to ER-α.

关 键 词：金雀黄素 子宫内膜癌 细胞增殖 雌激素受体 报告基因 

分 类 号：R285.5[医药卫生—中药学]