从息风药对肝癌细胞EMT的作用探讨"内风"是肝癌转移的关键病机

Metastasis is one of the most common causes of mortality among patients with hepatocellular carcinoma. More and more researchers are focusing on the problem of the prevention and treatment of hepatocellular carcinoma metastasis. Growing evidence indicates that the epithelial mesenchymal transition (EMT) confers on cancer cells precisely the set of traits, that is acquisition of the metastatic phenotype, that would empower them to disseminate from primary tumors and seed metastases, the strategy of impeding the process of EMT seems to be the central issue. In the theory of Traditional Chinese Medicine, the characteristic of the internal wind is consistent with the property of hepatocellular carcinoma metastasis. Our previous study have showed that several Xifeng drugs have the ability to inhibit the migrate of hepatoma cells in vitro. Therefor, we hypothesize that the internal wind is maybe the key pathogenesis in the metastasis of hepatocellular carcinoma, and the Xifeng drugs may inhibit the metastasis of hepatocellular carcinoma through impeding EMT process. In this study, we employ the technology of in vivo imaging, lentivirus vector transfection and siRNA et al, to observe the action of Xifeng drugs on the liver cancer metastasis, the peripheral tumor cells, and the hepatic cancer stem cell, to study the regulation of that on the expression of the EMT-associated protein in vitro and in vivo; and to investigate the mechanism of that on EMT from the regulation of transcription factor expression. Ultimately, we intend to clarify the action and molecular mechanism of Xifeng drugs on hepatocellular carcinoma metastasis. The study provide a new idea in the Traditional Chinese Medicine for the preventment of hepatocellular carcinoma metastasis, which has important theoretical and clinical value.

肝癌转移是导致肝癌患者死亡的主要原因之一，愈来愈多的研究者把目光聚焦于肝癌转移防治领域。研究显示肝癌转移的始动环节是上皮细胞-间充质转化（EMT），它使肝癌细胞获得运动能力及侵袭表型，基于逆转肝癌细胞EMT的策略成为该领域的热点之一。中医理论认为肝癌转移快、进展迅速与风邪善行数变特性一致，我们前期研究发现部分息风药具有抑制肝癌细胞体外迁移的作用。据此，我们提出“内风是肝癌转移的中医关键病机”,“息风药可能通过阻碍EMT过程预防肝癌转移”的假说。本研究从在体和体外两方面，应用活体成像、慢病毒转染和siRNA等技术，观察息风药对肝癌转移、外周血肝癌细胞数以及肝癌干细胞数的影响，检测其对肝癌细胞EMT标志蛋白的作用，进一步从对EMT转录因子表达的调控，探讨息风药对EMT的调节作用，以阐明息风药对肝癌转移的作用及分子机制，为中医药预防肝癌转移提供新的思路，具有重要的理论意义和临床价值。

肝癌转移是导致肝癌患者死亡的主要原因之一, 愈来愈多的研究者把目光聚焦于肝癌转移防治领域。 中医理论中“风邪善行数变、百病之长特性”特性与肝癌转移快、进展迅速的特点一致。. 为了阐明“内风是肝癌转移的关键病机之一”，本研究利用动物活体成像系统，在体观察了全蝎作用下裸鼠体内癌肿生长和转移的情况，并检测了发生转移的组织器官EMT相关标志蛋白的改变。接着采用血清药理学的研究方式，在离体水平分别从肝癌细胞的增殖活性、细胞的凋亡状态、细胞的迁移能力和细胞的侵袭水平四个方面，观察研究全蝎、僵蚕、全麻含药血清在体外对鼠源性肝癌细胞Hepa1-6产生的作用和影响。为深入了解潜在的作用机制，进一步以肝癌细胞Hepa1-6为研究对象，通过TGF-β1诱导建立了体外肝癌细胞EMT模型同时对其进行药物干预，对比观察了息风药作用后细胞EMT相关标志蛋白和转录因子Snail的表达变化。. 实验结果显示全蝎对肝癌转移裸鼠模型体内肿瘤的形成、生长和转移均表现出有效的抑制作用，并明显抑制了相关组织EMT的状态；息风药全蝎、僵蚕含药血清可有效抑制肝癌细胞Hepa1-6体外的增殖活性，诱导促进其凋亡，并明显降低细胞侵袭和迁移的能力；其作用可能通过抑制转录因子Snail的表达，对上皮细胞间充质转化的过程发挥负向调节作用，降低肝癌细胞的侵袭能力，从而抑制肝癌转移。. 本研究在中医治未病思想指导下,紧抓肝癌转移的始动环节,首次将“内风”与上皮细胞-间充质转化(EMT)相关联,从抑制肝癌细胞 EMT 这一重要靶点,研究息风药抗肝癌转移的作用机制。通过这一研究, 完善了肝癌转移的中医病机认识，为肝癌转移的中医治疗提出新思路,具有非常重要的理论意义和临床价值。