中草藥M1抗肝癌之研究

Abstract

肝細胞癌（Hepatocellular carcinoma）是目前人類常見的惡性癌症之一，據統計肝癌致死率二十年來是台灣癌症主要死因。主要致癌因子包含了肝炎病毒感染和其他原因如酒精濫用、黃麴毒素的攝取等。 目前以中草藥治療疾病機轉的研究獲得重視，本研究中以Huh7肝癌細胞株來檢測中草藥對於肝癌細胞的血管新生作用及轉移作用是否能有效抑制。結果，由10種中藥發現M1中草藥能夠以低於細胞生存率(Cell Viability)濃度的一半濃度以下有效抑制Huh7的細胞遷移能力(Migration)。中草藥M1 針對細胞膜上的受器酪胺酸激酶 (Receptor Tyrosine Kinase，RTK)中的表皮生長因子受體(Epidermal Growth Factor Receptor，EGFR)有效抑制其磷酸化，並且抑制其下游分子AKT (Protein Kinase B，PKB)、mTOR (mammalian target of rapamycin)、HIF1-α(Hypoxia-inducible factor 1-alpha) 和VEGF-A(Vascular Endothelial Growth Factor A)的表現。EGFR下游路徑蛋白 Ras (Rat sarcoma) 、 ERK (extracellular signal–regulated kinases)、轉錄因子Snail亦有被抑制表現。細胞轉移機制中，可分解細胞外基質(Extracellular Matrix，ECM)蛋白的酵素，基質金屬蛋白酶(Matrix Metalloproteinases，MMPs)中的MMP-2、MMP-9在細胞外活性和表現量能夠被有效的抑制和減少。Epithelial–mesenchymal transition (EMT) 中的指標蛋白，如: E-cadherin和N-cadherin表現量也有改變，抑制細胞的遷移能力。本研究也藉由免疫缺陷小鼠(NOD/SCID)建立腫瘤模式，並以口服M1來觀察是否可在活體(in vivo)內抑制腫瘤生長，結果在腫瘤生長大小、重量、蛋白表現上都被有效抑制，M1確實可在活體中抑制腫瘤生長。總結研究結果可，中草藥M1有其發展抗癌藥物的潛力。Hepatocellular Carcinoma (HCC) is one of higher malignant in cancers of human in the world. In addition, HCC mortality rate for 20 years is the leading cause of cancer death in Taiwan. The primary carcinogenic factors includes hepatitis virus infection and other are caused by alcohol abuse, and ingestion of carcinogen such as aflatoxin, etc. HCC has the characteristics of high level metastasis, angiogenesis, and recurrence after surgery. In this study, we focused on how to effectively prevent metastasis and angiogenesis of HCC. Recently, it becomes important to use the Chinese Herbal Medicine (CHM) as a therapeutic strategy for the treatment of diseases. Here, we found that among 10 kinds of CHMs, M1 could inhibit the cell migration at the dose less than half concentration of IC50 (the half maximal inhibitory of concentration). M1 significantly inhibited the expression of Receptor Tyrosine Kinase, Epidermal Growth Factor Receptor (EGFR), and downstream, Ras, ERK, and Snail. Furthermore, another downstream pathway protein, Protein Kinase B (AKT), mammalian target of rapamycin (mTOR), Hypoxia-inducible factor 1-alpha (HIF1-α), and Vascular Endothelial Growth Factor A (VEGFA) were also inhibited. And the Matrix Metalloproteinases (MMPs), including MMP-2, MMP-9, which play an important role in metastasis mechanism, were also inhibited by M1. Epithelial–mesenchymal transition (EMT) markers, such as E-cadherin and N-cadherin, were effective altered by M1 treatment. In the xenograft mice model, we observed the tumors growth sizes, weight, and protein expressions were effectively inhibited by M1. In conclusions, we demonstrate that M1 has an inhibitory effect to suppress angiogenesis and metastasis of HCC in vitro and in vivo.