芒柄花黄素通过MAPK/AKT通路诱导不同前列腺癌细胞凋亡的辩证研究

Prostate cancer is the first most common cancer among men in Western countries. Estrogen receptor play key roles in the development of prostate cancer. The isoflavone formononetin (C16H12O4) is a phytoestrogen and one of the main active components of red clover plants. Because of their structural similarity with estrogen, phytoestrogens can produce estrogenic or/and antiestrogenic effects. Our previous studies suggested that formononetin inactivated extracellular signal-regulated kinase1/2 (ERK1/2) mitogen-activated protein kinase (MAPK) signaling pathway in a dose-dependent manner, which resulted in increased the expression levels of BCL2-associated X (Bax) mRNA and protein, and induced apoptosis in LNCaP cells. At the same time, formononetin acted solely as estrogen agonists in prostate cells. In the present study, by cell counting, MTT assay, flow cytometry, Western blot, RT-PCR, siRNA, Xenograft tumor growth, immunohistochemistry, we identified that formononetin induced proliferation of prostate cells and apoptosis of prostate cancer cells via interrupting the cross-talk among multiple signaling pathways including MAPK and Akt. Furthermore, we focus on the effect of formononetin on the levels of p38/Akt protein. Our results provided the foundation for future development of formononetin for treatment of prostate cancer.

前列腺癌属于男性最常见的恶性肿瘤，雌激素受体(ER)对前列腺癌的发生与发展起到重要作用。芒柄花黄素是传统中药三叶草的主要活性成分之一，具有明显选择性雌激素受体调节剂作用。课题组研究表明芒柄花黄素能够与ERβ结合，作为拮抗剂通过ERK1/2 MAPK通路抑制前列腺癌细胞增殖，而预实验显示同一剂量时作为激动剂促进前列腺细胞增殖。本项目拟通过体外、体内模型，观察芒柄花黄素与雌激素受体各亚型结合情况及转录活性，检测其诱导前列腺癌细胞凋亡，阻滞癌细胞增殖周期但促进前列腺细胞增殖的作用，辩证的论证这些作用是否通过MAPK/Akt通路，分析两条通路的相互关系，并着重检测Akt/p38上下游基因蛋白表达情况，探讨芒柄花黄素对前列腺癌细胞及前列腺细胞不同的分子机制，以期探寻芒柄花黄素对不同靶细胞的个性化治疗。

前列腺癌属于男性最常见的恶性肿瘤，雌激素受体(ER)对前列腺癌的发生与发展起到重要作用。芒柄花黄素是传统中药三叶草的主要活性成分之一，具有明显选择性雌激素受体调节剂作用。课题组研究表明芒柄花黄素能够与ERβ 结合，作为拮抗剂通过ERK1/2 MAPK 通路抑制前列腺癌细胞增殖，而实验显示同一剂量时作为阻滞剂抑制前列腺细胞周期，同样的抑制细胞增殖。本项目通过体外、体内模型，通过MTT, 流式细胞术, Western blot, RT-PCR, siRNA , 免疫组化等不同技术手段，观察不同浓度芒柄花黄素诱导前列腺癌细胞凋亡，阻滞癌细胞增殖周期的作用，观察到这些作用通过MAPK/Akt 通路，干预IGF-1/IGF-1R，以及Bcl-2/Bax，P21WAF1, cyclinD1基因和蛋白表达，分析两条通路的相互关系，并着重检测P21WAF1, cyclinD1，Akt/p38 上下游基因蛋白表达情况，实验的数据可为众多的异黄酮类植物雌激素抗激素相关性肿瘤及促肿瘤生长提供新的思路与方向，以期为不同药物寻找各自特异性的作用靶点。