黃連於清胃散誘使口腔癌細胞株凋亡功效之研究

The study of induction of apoptosis in human oral cancer cell lines OC2 by Coptis Chinensis in Chingwaysan

廖翊傑(I-Chieh Liao)

指導教授 ： 黃裕峰

中山醫學大學/口腔醫學院/牙醫學系碩士班/碩士(2016年)

摘要

癌症早已成為國內十大死因排名第一位，其中口腔癌已躍昇為台灣男性癌症死亡率第四名，且為十大癌症罹患人數及死亡人數增加最快的癌症。目前，化學治療與外科手術、放射治療已成爲醫學界公認治療惡性腫瘤的三種主要方法，化學療法具有全身治療或輔助其他療法進而消滅亞臨床瘤細胞的效果，所以醫學界不斷地爲開發更有效的新型抗腫瘤藥物而努力。細胞凋亡，又稱爲程式性細胞死亡。一般而言是由生理或病理性因素誘發進而使細胞主動實施。細胞凋亡是維持細胞內環境穩定的自主而有序的死亡。目前雖已分離了一些與細胞凋亡有關的癌基因及抑癌基因，而對這些基因的生化機制及細胞凋亡在腫瘤發生、生長、退化中的地位還所知有限。不過，控制細胞凋亡已爲人們治療腫瘤提供了一條新思維。中醫藥雖有千年的歷史但對於腫瘤的治療往往流於不科學性及不確定性的偏方，因此以科學性的方法釐清中醫藥材的藥理機轉與建立使用上的理論基礎為推動中醫藥運用於腫瘤治療。本研究室先前以中藥方劑清胃散投藥於口腔癌細胞株，發現其可藉由細胞凋亡的路徑降低癌細胞的存活，清胃散是複方藥劑，其中含升麻、牡丹皮、黃連、當歸、生地黃等成分，本次以細胞存活率分析來探討清胃散中單方生藥和先前的清胃散複方各以不同濃度給藥後存活率的表現，用西方墨點法對p21及Bax蛋白分析，並觀察癌細胞去氧核醣核酸斷裂情形，發現清胃散各單方生藥中黃連會抑制癌細胞生長並提高p21和Bax蛋白量，也會使口腔癌細胞產生DNA片段化現象，證明黃連會使口腔癌細胞產生細胞凋亡抑制生長，其路徑可能藉由提高p21或Bax蛋白量誘發細胞凋亡，與先前的清胃散實驗結果相似，可知單方黃連於清胃散經細胞凋亡而抑制口腔癌細胞生長效果之主要角色。

關鍵字

黃連 ； 清胃散 ； 口腔癌 ； 細胞凋亡

並列摘要

Cancer has ranked number one among the top ten leading causes of death in Taiwan. Among all cancers, oral cancer has risen to the fourth highest cause of cancer-related mortality among men in Taiwan. Moreover, it is also the cancer that has had the most dramatic change in the number of diagnosed patients and the number of total deaths among the top ten leading cancers. Currently, chemotherapy, surgery, and radiation therapy have been well acknowledged to be the three main treatments for malignant tumors in the medical field. Chemotherapy has effects on making cancer cells extinct clinically by systematic treatment or as an adjuvant to other therapies. Thus, there have been constant efforts to develop new anti-tumor drugs with better therapeutic effects in the medical field. Apoptosis, also called programmed cell death, is a spontaneous and sequential death controlled by genes to maintain stability within the cellular environment. Although some oncogenes and cancer suppressor genes related with apoptosis have been identified recently, there is limited understanding of the biochemical mechanism of the genes and the roles of apoptosis on tumorigenesis, tumor development, and tumor regression. However, modulating apoptosis has shed new light on tumor treatment. Even though Chinese Medicine has a history spanning several thousand years, its treatment of tumors has been considered to be a non-scientific and uncertain folk treatment. Thus, in order to promote tumor treatment using Chinese Medicine, it is necessary to understand the pharmacological mechanisms of Chinese herbs using scientific methods and to build a theoretical basis for their use. We have previously found that the survival rate of oral cancer cell line (OC2) decreases via apoptosis pathway when treated with a Chinese medicinal formula, chingwaysan. In this study, we further investigated the effect of different Chinese herbs of chingwaysan at various concentrations on the cell survival rate. By analyzing p21 and BAX (pro-apoptotic protein) and evaluating the DNA breakage in cancer cells, we found that coptis chinensis inhibits cancer cell growth and increases the amount of anti-cancer proteins, resulting in DNA fragmentation. The results are similar to the results of the previous experiments with chingwaysan. Therefore, coptis chinensis plays an important role in the inhibiting effects of chingwaysan on oral cancer cell growth via apoptosis.

並列關鍵字

Coptis Chinensis ； Chingwaysan ； apoptosis ； oral cancer

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