研究天然粗萃物對肺腺癌和抗藥性肺腺癌細胞株的抑癌作用

Study the Anti-cancer effects of Natural Plant Extracts in vincristine drug resistance lung adenocarcinomas

郭珈夆(Chia-Feng Kuo)

指導教授 ： 蔡女滿

中山醫學大學/醫學科技學院/醫學檢驗暨生物技術學系/碩士(2019年)

https://doi.org/10.6834/CSMU201900257

摘要

肺癌 (Lung cancer) 為目前全世界發生率及死亡率最高的癌症，在台灣近十年也都排名第一名，其中又以肺腺癌 (Adenocarcinoma)佔最多，平均五年存活率只有19.4%，尤其到晚期更下降至5.2%，目前晚期治療方法以化療或輔助性療法為主，但病患預後不佳，甚至產生抗藥性，因此尋找具有潛力能解決臨床上抗藥性瓶頸之藥物是重要的。本篇論文研究目的為探討天然粗萃物是否對肺腺癌及抗藥性肺腺癌細胞有抑癌作用。首先先利用MTT assay建立篩藥平台，確定抗藥性肺腺癌細胞A549/V16確實具有抗藥性，並使用五種天然粗萃物廣藿香 (Pogostemon cablin, PPa)、雪松 (Cedrus atlantica, CAt)、滇藏方枝柏 (Juniperus indica Bertol, JIB)及純化物Patchouli alcohol (PAT)及Cedrol，篩選出相對於臨床藥物長春新鹼 (Vincristine)對抗藥性肺腺癌細胞A549/V16較為有效之純化物PAT，之後實驗結果顯示PAT可使細胞週期停滯於G0/G1 phase，另外Sub-G1百分比也有增加之趨勢，接著利用TUNEL assay觀察到細胞凋亡的相關型態細胞染色質濃縮、DNA斷裂及細胞凋亡小體，而後利用Immunocytochemistry (ICC) 染色法和西方點墨法觀察到PAT可誘發細胞凋亡之外在與內在途徑使細胞進行細胞凋亡。綜合以上結果，PAT在對於抗藥性肺腺癌治療上具有高度開發的潛力。

關鍵字

肺腺癌 ； 抗藥性 ； 長春新鹼 ； Patchouli alcohol (PAT) ； Cedrol

並列摘要

Lung cancer is the most common cause of cancer death around the world and in Taiwan, and the adenocarcinoma are the most type of lung cancer cases. The five-year survival rate of patients with adenocarcinoma is approximately 19.4%, especially the distant stage is less to 5.2%. The treatment for the distant stage are mainly use chemotherapy and palliative intent. However, more patients with advanced lung cancer have a poor efficacy because of the multi-drug resistance. Therefore, it is important to develop a potential drug for the drug resistance lung adenocarcinoma. The aim of this study is to investigate the anti-cancer activity and mechanism in lung adenocarcinoma cell and vincristine resistance lung adenocarcinoma cell line. At the first, we used MTT assay to determine that vincristine resistance lung adenocarcinoma cell (A549/V16) had higher IC50 value than lung adenocarcinoma cell after treating vincristine. Then, we used the extracts of Pogostemon cablin (PPa), Cedrus atlantica (CAt), Juniperus indica Bertol (JIB) and Patchouli alcohol (PAT) and Cedrol pure compounds. The results showed that PAT and Cedrol had higher cytotoxicity in lung adenocarcinoma cell A549 and vincristine resistance cell A549/V16. In addition, PAT could induce cell cycle arrest at G0/G1 phase and increase percentage of Sub-G1 phase. The cell death and apoptosis morphology was observed by TUNEL assay. Then, the extrinsic and intrinsic apoptosis associated proteins were detected by Immunocytochemistry (ICC) stain and western blot. The results showed that PAT could induced cell apoptosis. Taken together, PAT have potential develop for the treatment of drug-resistant lung adenocarcinoma.

並列關鍵字

Lung adenocarcinoma ； Drug-resistant ； Patchouli alcohol (PAT) ； Cedrol

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