大豆染料木黄酮与内源性雌二醇双向生物学活性及分子机制的研究

Genistein is one of the main anti-cancer factor of the soy isoflavone family, the combined effect of estrogen as a class and human endogenous sex hormones on breast cancer in women is to determine the key points to prevent and control the effects of breast cancer. Epidemiology and in vitro experiments showed that premenopausal soy isoflavone intake and breast cancer risk was a significant negative correlation, but it may stimulate the growth of breast cancer in postmenopausal showed that endogenous estrogen levels its prevention and treatment effect. The existing reports are the genistein monomer to study the role of breast cancer cells, while ignoring the interaction with the endogenous estrogen in the human body and its different and even opposite results, In addition, different types of estrogen receptor and the biological effects of genistein, there is a big difference. The debate on soy isoflavones and breast cancer relationship is even more acute, the lack of strong enough to reach a common supporting evidence. This study intends to genistein, the endogenous estrogen and different estrogen receptor types combine the three genome-wide cDNA microarray analysis as a high-throughput gene expression analysis tools to obtain the expression of differences under the different approach genetic profile, and explore related genes.From the micro-level, and based on an overall perspective and clear genistein bidirectional molecular mechanisms of biological activity and its metabolic pathway, and provide a scientific basis for genistein breast cancer prevention.

染料木黄酮是大豆异黄酮家族的主要抗癌因子之一，其作为类雌激素与人体内源性激素对女性乳腺癌的共同影响是决定其防控乳腺癌作用效果的关键点。流行病学及体外实验表明，绝经期前大豆异黄酮摄入量与乳腺癌发病风险呈明显的负相关，但在绝经后则有可能会刺激乳腺癌的增长，表明内源性雌激素水平不同其防治效果不同。现有报导均以染料木黄酮单体来研究对乳腺癌细胞的作用，而忽略人体内内源性雌激素与其的交互作用，得出不同甚至相反的结果，此外，雌激素受体类型不同，染料木黄酮生物学效应存在较大差别。对大豆异黄酮和乳腺癌关系的争论更加尖锐，缺乏强有力的、足以达成共性的支持证据。.本研究拟将染料木黄酮、内源性雌激素与不同雌激素受体类型三者有机结合，以全基因组表达谱芯片作为一种高通量的基因表达分析工具，获取不同处理方式下差异表达基因谱，探索相关基因群，从微观层次，并立足于整体角度明确染料木黄酮双向生物学活性的分子机制及其代谢通路。

因内源性雌激素水平不同、雌激素受体类型不同及染料木黄酮与雌激素相互作用等因素，染料木黄酮对乳腺癌的生物学效应不尽相同甚至完全相反，然而其机制并不明确。故本研究选取ER类型不同的乳腺癌细胞MDA-MB-231（ERβ）和T47D（ERα+ERβ）作为模型，开展了3方面的工作：首先采用MTT和流式技术分析染料木黄酮在不同雌二醇水平下对不同ER类型的乳腺癌细胞表型的影响；其次采用全基因组表达芯片技术检测染料木黄酮在不同雌二醇水平下影响乳腺癌细胞表型的差异表达基因；最后经过生物信息学分析及相关实验验证具体参与染料木黄酮介导的乳腺癌细胞表型调控相关基因。通过该项目的研究取得了以下结果：一是证实了染料木黄酮在乳腺癌细胞中功能的发挥不仅与雌二醇水平相关而且与ER受体类型也有关系。在不同雌二醇水平下，染料木黄酮引起乳腺癌细胞发生周期阻滞、抑制细胞增殖和诱导细胞凋亡的能力均不同。二是初步明确了染料木黄酮在乳腺癌中发挥调控功能的相关分子及信号通路。全基因组表达谱分析结果获得了一系列与细胞增殖、生长、凋亡有关的差异基因。后期采用实时荧光定量PCR技术验证了这些差异表达基因。这些研究结果将为后续深入探讨染料木黄酮在乳腺癌中发挥功能的分子机制提供坚实的基础，为临床乳腺癌治疗提供新的干预靶点及理论依据。