【作者】 刘念周；

【导师】 陈锋；

【作者基本信息】 南华大学 ， 卫生毒理学， 2005， 硕士

【摘要】 目的: 通过研究己烯雌酚对体外培养的雌激素受体阳性人乳腺癌MCF-7细胞中的WISP-2基因表达的影响,以雌二醇作对照,探讨己烯雌酚在乳癌发生中的作用;研究MCF-7细胞株WISP-2基因对BCL-2基因的影响,探讨两个基因在抑制癌细胞凋亡、促进癌细胞增殖过程中的作用。 方法: 采用MTT法检测己烯雌酚对MCF-7细胞生长活性的影响。运用免疫细胞化学方法和逆转录-聚合酶链反应方法检测己烯雌酚对体外培养的人乳腺癌细胞MCF-7中WISP-2mRNA和WISP-2蛋白表达影响,并以雌二醇作阳性对照组。运用免疫细胞化学方法检测人乳癌细胞MCF-7中WISP-2蛋白表达变化时BCL-2的蛋白表达状况。 结果: 1)己烯雌酚使MCF-7增殖率增加。 2)己烯雌酚作用于MCF-7细胞使WISP-2mRNA和WISP-2蛋白表达上升,高浓度的己烯雌酚的效应高于低浓度的己烯雌酚的效应。WISP-2mRNA和蛋白表达随己烯雌酚作用时间的延长而增加,其效应与雌二醇类似。 3)用反义核苷酸抑制乳癌细胞MCF-7中WISP-2蛋白的表达后,BCL-2蛋白表达降低。 结论: 1) 己烯雌酚能促进MCF-7细胞增殖。 2) 己烯雌酚能诱导MCF-7细胞中的WISP-2基因表达增加,作用强度随更多还原

【Abstract】 Object:l)To explore the effects of diethylstilbestrol on the expression of the WISP-2 gene in human breast cancer cell line MCF-7,and to compare its effect with 17- β estradiol. The research is to study the role of diethylstilbestrol in the development of human breast cancer, and its mechanism.2)To explore the effects of WISP-2 gene on the expression of the BCL-2 gene in human breast cancer cell line MCF-7. The research is to study the role of them in the inhibition of cell apoptosis and acceleration of cell proliferation .Methods:MCF-7 cell were cultured in vitro, and exposed to various concentrations of diethylstilbestrol in the different time, the expression of WISP-2 mRNA and its protein were measured using RT-PCR and immunohistochemistry approach. The experiment set up vacuity negative control , and 17-β estradiol positive control, the expression of BCL-2 protein was measured using immunohistochemistry approach when the expression of WISP-2 protein changed in the MCF-7 cell. To detect the proliferation was using MTT approach.Results:1) Diethylstilbestrol accelerated the MCF-7 cell proliferation.2) Exposed to diethylstilbestrol, expression of WISP-2 mRNA and its protein was increased. The effect of high concentration diethylstilbestrol was更多还原