基于PI3K/AKT/mTOR信号转导通路探讨西黄丸干预乳腺癌前病变的作用机制

PI3K/AKT/mTOR signal transduction pathway is the most closely related to the development of breast cancer. Preliminary studies have confirmed that the Xihuang Pill has a good therapeutic effect on breast precancerous lesions. It can significantly reduce the expression levels of the p-AKT, p-mTOR and VEGF proteins of the PI3K/AKT/mTOR signal transduction pathway, which suggesting that good effect of Xihuang Pill is closely related to this pathway. For further research, we establish the rat breast precancerous lesion disease models by DMBA combined with estrogen and progesterone sequential method and detect the proteins level of p-PI3K, p-AKT, p-mTOR, PTEN, p-TSC2, p-S6K1, p-4E-BP1, VEGF and genes expression of PTEN and VEGF by Western-Blot, RT-PCR and LC-MS techniques. Analysis impact of Xihuang Pill on key signaling molecules of this pathway and confirmed it via MCF-10AT cell model. To study the intervention mechanisms of Xihuang Pill on breast precancerous lesions through the PI3K/AKT/mTOR signaling pathway, reveal the mechanisms for treatment and prevention or blocking the development of breast cancer.

PI3K/AKT/mTOR信号通路与乳腺癌发生发展最密切相关。西黄丸是治疗乳腺癌前病变、防治乳腺癌之经典名方，前期研究其可显著降低病变细胞该信号通路蛋白p-AKT、p-mTOR和VEGF的表达，提示其良好疗效与其干预该信号通路密切相关。本项目采用Western-Blot、RT-PCR和LC-MS等技术，以DMBA联合雌孕激素序贯诱导动物模型，通过检测该通路分子PI3K、AKT、mTOR磷酸化水平，负性调节因子PTEN、p-TSC2及通路下游p-S6K1、p-4E-BP1、VEGF蛋白及基因表达变化水平，明晰西黄丸作用于该通路的可能关键信号分子；并利用MCF-10AT细胞模型确证西黄丸通过可能关键信号分子发挥防治作用，以研究西黄丸对乳腺癌前病变细胞该信号通路的干预机制，揭示其有效治疗乳腺癌前病变和预防、阻断乳腺癌发生发展的作用机理及作用靶点。为西黄丸有效防治乳腺癌提供现代分子生物学依据。

PI3K/AKT/mTOR信号通路与乳腺癌发生发展最密切相关。西黄丸是治疗乳腺癌前病变、防治乳腺癌之经典名方，但分子机制不明，前期研究其可显著降低病变细胞该信号通路蛋白p-AKT、p-mTOR和VEGF的表达，提示其良好疗效与其干预该信号通路密切相关。本项目采用Western-Blot、RT-PCR和免疫组化等技术，以DMBA联合雌孕激素序贯诱导动物模型，通过检测该通路分子PI3K、AKT、mTOR磷酸化水平，负性调节因子PTEN、p-TSC2及通路下游p-S6K1、p-4E-BP1、VEGF蛋白及基因表达变化水平，明晰了西黄丸通过PI3K/AKT/mTOR信号通路实现调控乳腺癌前病变细胞周期，抑制细胞生长增值，诱导细胞凋亡，并利用乳腺癌前病变细胞模型MCF-10AT进行了验证，揭示了其有效治疗乳腺癌前病变和预防、阻断乳腺癌发生发展的作用机理及作用靶点。为西黄丸有效防治乳腺癌提供现代分子生物学依据。