论文部分内容阅读
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 effect of genistein on the proliferation of estrogen and progesterone receptor negative cell line (HEC-1B) in endometrial carcinoma and the mechanism of its receptor, so as to provide a theoretical basis for the anti-tumor effect of genistein. Methods In vitro culture of endometrial carcinoma HEC-1B cells, the concentrations of genistein were 10 × 10-6, 20 × 10-6, 40 × 10-6, 80 × 10-6, 160 × 10-6 mol / L, the action time were 24,48,72,96 h. The effects of genistein on proliferation of endometrial cancer cells were detected by MTT assay. Meanwhile, luciferase reporter gene was used to detect the effects of genistein on estrogen response ( ERE) on estrogen receptor alpha (ER-alpha) and estrogen receptor beta (ER-beta) reporter gene expression, and to compare their differences in the role of ER alpha and ER beta. Results Genistein had a significant inhibitory effect on the proliferation of HEC-1B cells in vitro (P <0.01), and its inhibitory effect increased gradually with the increase of the concentration of the drug and the prolongation of the action time, in a dose and time dependent manner Sex. The expression of reporter gene was significantly increased in genistein at a concentration of 20 × 10-6 and 40 × 10-6mol / L compared with that of the blank control group (P <0.01), and in a concentration-dependent manner The level of genistein up-regulated by ER-β was higher than that of ER-α, the difference was statistically significant (P <0.01). Conclusion Genistein can inhibit the proliferation of endometrial cancer cells. This inhibition may be through regulating the expression of ER-α and ER-β in endometrial carcinoma cells and adjusting the ER-β / ERα-ratio.