三黄煎剂基于抗慢性氧化应激介导 PI3K通路抑制乳腺癌微环境侵袭的实验研究

Based on the primary effects of San Huang Jian Ji on the inhibition of MCF-7 cell proliferation, anti-angiogenesis, anti-inflammation，the project explore the effect of it on the inhibition of MCF-7 cell chronic oxidative stress and the mechanism of it on the inhibition of invasion and metastasis of breast cancer. Based on anti-chronic oxidative stress by activating the pathway of Nrf-2/ARE mediating PI3K pathway, which can provide a method of finding the mechanism and target spot with the interference of San Huang Jian Ji inhibiting invasion and metastasis of breast cancer. Through establishing the breast cancer nude model in the chronic oxidative stress microenvironment, the effect and mechanism of regulating chronic oxidative stress and inhibiting invasion and metastasis with the interference of San Huang Jian Ji are further confirmed. The project provides the experiment evidence that Traditional Chinese Medicine theory about Ying-yang balance applies to explain the phenomenon of regulating the balance of oxidative stress in order to control invasion and metastasis of breast cancer.

本研究在三黄煎剂基于抗氧化应激抑制MCF-7细胞增殖侵袭、血管新生、炎症反应的前期实验与临床基础上，进一步探查其对抑制MCF-7细胞慢性氧化应激微环境的影响及探讨其抑制肿瘤侵袭转移的机制。研究基于核因子Nrf-2/ARE通路抗慢性氧化应激介导PI3K通路，寻找出三黄煎剂抑制肿瘤微环境侵袭转移的机制与靶点，并经在体实验证实三黄煎剂调控慢性氧化应激抗肿瘤侵袭转移的功效与机制。为中医药阴阳平衡理论指导调控机体氧化应激平衡控制肿瘤侵袭转移提供实验依据。

氧化应激可激活PI3K/AKT通路促进乳腺癌细胞增殖、血管新生、侵袭及转移，也可引起乳腺癌内分泌药物的耐药。本研究在三黄煎剂基于抗氧化应激抑制MCF-7细胞增殖、侵袭、血管新生、炎症反应的前期实验与临床基础上，进一步探讨三黄煎剂干预PI3K/AKT通路调控慢性氧化应激微环境下乳腺癌的侵袭和转移。.主要研究内容：建立慢性氧化应激MCF-7细胞模型；探讨三黄煎剂对慢性氧化应激MCF-7细胞的增殖、凋亡、侵袭、转移及PI3K/AKT通路的作用，加入PI3K/AKT信号通路的抑制剂LY294002和激动剂IGF-1后，进一步证实三黄煎剂对PI3K/AKT信号通路的影响。构建稳转的上调MCF-7/shARE-Nrf2和下调MCF-7/siNrf-2细胞系，裸鼠接种MCF-7细胞及MCF-7/siNrf-2细胞，观察三黄煎剂对肿瘤的生长、侵袭、转移及PI3K/AKT通路的影响。.重要结果：成功的建立了慢性氧化应激MCF-7细胞模型；三黄煎剂能够呈浓度依赖性的抑制慢性氧化应激MCF-7细胞的增殖，诱导凋亡，抑制黏附、迁移、侵袭及PI3K/AKT通路；加用PI3K/AKT信号通路的抑制剂LY294002和激动剂IGF-1，进一步证实了三黄煎剂以PI3K/AKT为靶点，调控慢性氧化应激MCF-7细胞的增殖和转移。成功的建立了构建稳转的上调MCF-7/shARE-Nrf2和下调MCF-7/siNrf-2细胞系模型。裸鼠接种MCF-7细胞及MCF-7/siNrf-2细胞成瘤，三黄煎剂干预后可明显抑制肿瘤的生长、提高抗氧化应激能力、抑制了肿瘤的侵袭和转移，且可抑制PI3K/AKT水平。.关键数据：发表中文核心期刊论文1篇，SCI论文1篇在返修阶段，获江苏省中医院科学技术进步奖一等奖，培养博士研究生1人，培养硕士研究生1人，参加国内会议交流2次。.科学意义：三黄煎剂基于调控PI3K/AKT通路抑制慢性氧化应激微环境下乳腺癌的增殖、侵袭和转移，有效预防乳腺癌的复发和转移，因此，有广阔的临床运用前景。