基于miR-7/FAK途径探讨西黄丸抑制结肠癌细胞上皮-间质转化（EMT）的机制

Epithelial-mesenchymal transiton (EMT) is a key process in the metastasis of colon carcinoma . Xihuang Pill is an effective drug for the prevention of colon cancer metastasis. In our preliminary studies, there are definitive evidence that inhibition of the metastasis of cancer by Xihuang Pills is closely related to the inhibition of EMT. In our preliminary studies, we found that Xihuang Pills can up-regulate miR-7 expression bu down-regulate FAK expression. FAK play an important role in EMT in colon carcinoma cells. As a direct regulator of FAK expression, miR-7 can inhibit EMT in colon carcinoma. Based on the above evidence, we hypothesize that miR-7/FAK pathway may play an important role in the prevention of EMT by Xihuang pills.. Based on the preliminary studies, we will investigate the role of miR-7/FAK pathway in prevention of EMT in colon carcinoma cells by Xihuang Pills using shRNA technology to knockdown miR-7 in colorectal cancer cell lines and mice. The project aims to reveal the mechanisms of preventing colorectal carcinoma from EMT in colon carcinoma cells by Xihuang Pills, and to provide evidence for prevention of metastasis in colon carcinoma by traditional Chinese medicine.

上皮-间皮转化（EMT）是结肠癌细胞转移的关键机制。在临床中，西黄丸防治结肠癌转移疗效确切。本课题组前期研究表明西黄丸抑制结肠癌转移效应同其抑制结肠癌细胞EMT有关，进一步研究发现西黄丸可以上调结肠癌细胞miR-7（具有抑制EMT作用的microRNA）的表达，下调结肠癌细胞FAK（miR-7的靶基因）的表达。基于上述证据，我们提出假说：miR-7/FAK途径可能在西黄丸调控结肠癌细胞EMT中发挥重要作用。本研究拟在前期工作的基础上，采用分子生物学技术，构建慢病毒载体、建立稳定表达细胞株和构建裸鼠皮下移植瘤模型，在细胞和动物水平分别沉默miR-7的表达或上调FAK的表达，进一步观察miR-7/FAK途径在西黄丸抑制结肠癌细胞EMT中的作用及对结肠癌细胞转移的影响。本项目旨在揭示西黄丸抑制结肠癌细胞EMT的作用与机制，为中医药防治结肠癌转移提供科学证据。

上皮-间皮转化（EMT）是结肠癌细胞转移的关键机制。在临床中，西黄丸防治结肠癌转移疗效确切。本项目前期研究表明西黄丸抑制结肠癌转移效应同其抑制结肠癌细胞EMT有关，进一步研究发现西黄丸可以上调结肠癌细胞miR-7（具有抑制EMT作用的microRNA）的表达，下调结肠癌细胞FAK（miR-7的靶基因）的表达。本项目在前期工作的基础上，采用分子生物学技术，构建慢病毒载体、建立稳定表达细胞株和构建裸鼠皮下移植瘤模型，在细胞和动物水平分别沉默miR-7的表达或上调FAK的表达，进一步观察miR-7/FAK途径在西黄丸抑制结肠癌细胞EMT中的作用及对结肠癌细胞转移的影响。本项目结果显示：①西黄丸可以抑制结直肠癌细胞体外增殖能力，促进细胞凋亡，并抑制细胞体外迁移及侵袭能力；在体内实验中，发现西黄丸可以抑制结直肠癌皮下移植瘤的生长和转移。②沉默miR-7后，西黄丸体内外抗癌效应下降，且EMT标志物升高；③过表达PARP-1表达，西黄丸体内外抗癌效应下降，且EMT标志物升高；④在临床研究中，发现西黄丸可以减轻放射性直肠炎的程度，提高患者生活质量。本项目揭示西黄丸抑制结肠癌细胞EMT的作用与机制之一，为中医药防治结肠癌转移提供科学证据。