探討廣藿香植物萃取液對於肝癌之生物活性影響

A study of anticancer effect and mechanism of Pogostemon cablin extract in hepatocellular carcinoma

黃曉凡(Xiao-Fan Huang)

指導教授 ： 許國堂

共同指導教授 ： 蔡女滿(Nu-Man Tsai)

中山醫學大學/醫學院/醫學研究所/博士(2021年)

https://doi.org/10.6834/csmu202100023

摘要

肝癌為台灣及全球癌症致死率的前 3 名，其中肝細胞癌 (Hepatocellular carcinoma, HCC)佔肝癌發生率的60～70%。臨床檢測出肝癌多為中晚期並伴隨著肝功能異常、患者對化療藥物反應率低和對標靶藥物容易產生抗藥性。目前科學家仍不斷的開發新藥或治療模式來治療HCC，因而積極開發低毒性且能治療或預防HCC的復發或轉移之藥物為迫切性的需求。於本實驗室先前的研究結果中，透過一系列植物萃取液及以生物活性導向之篩藥平台，篩選出對HCC細胞有明顯抑制效果之植物粗萃物為廣藿香粗萃物 (Pogostemon cablin exrtact, PPa extract)。近期科學性文獻中，已證實廣藿香具有抗菌、抗發炎及抑制大腸癌及肺癌之生長等作用，但其詳細抑癌機制尚未被深入研究。因此，本研究之目的為探討PPa extract對抗HCC的抑癌的作用，研究設計分為五個階段：(1) 篩選有效抑制HCC細胞生長之植物粗萃物並研究其機制；(2) 探討PPa extract對HCC之抗癌作用及鑑定PPa extract之有效成分；(3) 探討有效純化物對HCC之抗癌作用；(4) 探討有效純化物合併Sorafenib對HCC之抗癌作用；(5) 研究有效純化物對HCC細胞轉移之影響及其機制。總結上述，透過此實驗設計驗證PPa extract能夠透過誘發大量活性氧化物 (Reactive oxygen species, ROS)促使DNA損傷，進而使細胞週期停滯於G0/G1 phase和啟動外在 (FAS/FASL)及內在 (Bax/Bcl-2)細胞凋亡路徑來抑制HCC細胞生長。此外，PPa extract與Sorafenib具有協同作用，可通過AKT/mTOR途徑有效抑制HCC細胞增殖並減少HCC細胞的再生。在動物模型中，PPa extract也可抑制VEGF/VEGFR之蛋白表達並誘導細胞凋亡來抑制HCC腫瘤生長並延長動物壽命，而且PPa extract在體內幾乎沒有觀察到明顯的生理及病理毒性。最終，期望能將PPa extract或其純化物繼續朝向化學預防之防癌保健商品或是臨床用於治療HCC的新一代抗癌藥物，或者作為化療佐劑與臨床藥物搭配使用。

關鍵字

肝細胞癌 ； 廣藿香 ； 細胞凋亡 ； 細胞週期 ； 蕾莎瓦膜衣錠 ； 協同作用 ； 轉移

並列摘要

Liver cancer is estimated top three cancer mortality in Taiwan and the world. Hepatocellular carcinoma (HCC) accounts for 60～70% of the incidence of liver cancer. After diagnosed, the patients with advanced liver cancer account for more than half and accompany by abnormal liver function. Besides, patients have a low response rate and strong side effects to chemotherapy drugs and are easily resistant to target therapy drugs. Until now, scientists still constantly develop new drugs or treatment modalities to achieve more effective treatment for liver cancer. Therefore, it is great need to develop a drug with fewer side effects, curative and can prevent recurrence of liver cancer. In our previously results which was tested by drugs screening platform with biological activity guide revealed that Pogostemon cablin extract (PPa extract) inhibited HCC cells growth and has been confirmed its’ antibacterial activity, antiinflammatory activity and antitumor capacity on colorectal cancer and lung cancer. However, its detailed mechanism has not yet been extensively studied in HCC. As a result, the purpose of this study is to explore the anti-cancer effect of PPa extract against HCC. The study design is divided into five directions: (1) Screening the plant crude extracts that inhibit the growth of hepatoma cells; (2) Studying the anti-tumor effects and mechanisms of the extract and identifying the effective components of the extract; (3) Exploring the anti-cancer effect of effective components on HCC; (4) Investigating the anticancer effect of effective components combined with sorafenib on HCC;(5) Studying the effects of effective components on the metastasis mechanisms on HCC. In summary, PPa extract can trigger production reactive oxygen species (ROS) to damage DNA resulting in cell cycle arrest at G0/G1 phase and activating of extrinsic (FAS/FASL) and intrinsic (Bax/Bcl-2) apoptosis pathway to inhibit HCC cell growth. In addition, PPa extract and sorafenib have a synergistic effect on inhibition of HCC proliferation, reduction of AKT/mTOR signaling and suppression of HCC cell regrowth. In animal models, PPa extract can suppress HCC tumor growth and prolong the lifespan by inhibiting the protein expression of PCNA and VEGF/VEGFR and inducing apoptosis. Furthermore, PPa extract has little obvious physiological and pathological toxicity in vivo. Ultimately, we hope that PPa extract and the effective components can develop as health care products, a new generation of anti-cancer drugs or as adjuvant for chemotherapy.

並列關鍵字

Hepatocell u lar carcinoma ； Pogostemon cablin extract ； Apoptosis, ； Cell cycle ； Synergistic effect ； Sorafenib ； Metastasis

參考文獻

1. Mak, L. Y.; Cruz-Ramon, V.; Chinchilla-Lopez, P.; Torres, H. A.; LoConte, N. K.; Rice, J. P.; Foxhall, L. E.; Sturgis, E. M.; Merrill, J. K.; Bailey, H. H.; Mendez-Sanchez, N.; Yuen, M. F.; Hwang, J. P., Global epidemiology, prevention, and management of hepatocellular carcinoma. Am Soc Clin Oncol Educ Book 2018, 38, 262-279, 10.1200/EDBK_200939.

2. Venook, A. P.; Papandreou, C.; Furuse, J.; de Guevara, L. L., The incidence and epidemiology of hepatocellular carcinoma: a global and regional perspective. Oncologist 2010, 15 Suppl 4, 5-13, 10.1634/theoncologist.2010-S4-05.

3. Yim, H. J.; Suh, S. J.; Um, S. H., Current management of hepatocellular carcinoma: an Eastern perspective. World J. Gastroenterol. 2015, 21, 3826-3842, 10.3748/wjg.v21.i13.3826.

4. Dhanasekaran, R.; Bandoh, S.; Roberts, L. R., Molecular pathogenesis of hepatocellular carcinoma and impact of therapeutic advances. F1000Research 2016, 5, 10.12688/f1000research.6946.1.

5. Lencioni, R., Chemoembolization in patients with hepatocellular carcinoma. Liver cancer 2012, 1, 41-50, 10.1159/000339019.