新型組合藥物抑制口腔癌細胞生長之研究

Studies on a novel drug combination for inhibition of oral cancer cell growth

李智蓉(Chih-Jung Lee)

指導教授 ： 李宣信 ； 蕭水銀

中山醫學大學/健康管理學院/公共衛生學系碩士班/碩士(2017年)

摘要

台灣目前口腔癌發生率及死亡率日漸嚴重，根據衛生福利部統計，2015年為十大癌症死亡率第五名，在台灣口腔癌約佔所有癌症之6.7%，是男性所罹患的主要癌症中發生率和死亡率增加最快者，且男性死亡率為女性之11.1倍，也是國人男性所有癌症死亡年齡中位數最年輕的，約為58歲，比所有癌症死亡年齡中位數年輕約10歲。
產生口腔癌之因素很多，包括致癌物及其引發的發炎、氧化壓力及自由基等等，均會造成DNA損傷，引發細胞癌化，口腔癌的發生與口腔長期受到刺激，如嚼食檳榔等有關，根據統計，台灣約九成口腔癌患者都有嚼食檳榔的習慣。
目前口腔癌治療有手術切除，放射治療及化學治療等，最近幾年，由於藥物的進步以及強調保留器官功能的前提下，很多患者不經手術，直接接受放射線治療同步合併做化學治療，然而常用的化學藥物如5-FU (5-fluoro-uracil) 、 cisplatin (順鉑) 、 paclitaxel (taxol；太平洋紫杉醇) 、 docetaxel (taxotere；歐洲紫杉醇)、gemcitabine (gemzar；健擇) 、 vinorelbine (navelbine；溫諾平) 、 capecitabine (xeloda；截瘤達)雖具有優越的療效，但也伴隨明顯副作用，如噁心、嘔吐、骨髓抑制(免疫力下降)及抗藥性等。因此，研究具有較好療效且副作用少的新藥物，乃是目前急待解決的問題。
本研究選擇不同作用機制的藥物，擬探求新型組合藥方，希望不同藥物組成，可互相加強藥效，而降低藥物使用的劑量，進而預期降低副作用。最近研究顯示，發炎、氧化跟細胞癌化有密切關係，另外，谷氨酸(Glutamate)受體活化可促進腫瘤細胞的生長，因此，本研究選擇使用了具有抗氧化、抗發炎及保護細胞的薑黃素(C)，並將和具有拮抗谷氨酸受體活化的Memantine (M)，及目前常使用的癌症化學治療藥物太平洋紫杉醇Taxol (T)及艾黴素Doxorubicin (D)組合，利用細胞存活率分析(MTT assay)研究這些組合藥方對人類口腔鱗狀細胞上皮癌細胞株(oral squamous cell carcinoma cell line, OECM-1)及人類牙齦上皮細胞株(Smulow-Glickman gingival cells, SG cell)生長的影響，並計算combination index (CI)及potency ratio (P.R.)以闡明組合藥物抑制口腔癌細胞OECM-1生長效果是否比單用更佳，以及和正常牙齦上皮細胞SG相比，組合藥物抑制癌細胞生長有效濃度是否較低。
本研究發現抗癌藥物對於口腔癌細胞生長抑制作用，作用長時間的效果較佳，TC組合及TCM組合對於口腔癌細胞生長的抑制比單用及SG效果較佳，在藥物作用21小時Taxol+Curcumin組合CI=0.73，P.R.為1.4，在藥物作用43小時Taxol+Curcumin+Memantine 組合CI=0.64，為增益作用，有互相加強療效，將可預期作臨床試驗，進一步證明此併用之組成藥物，可能提高抗癌效果，降低人類口腔癌治療藥物之副作用，如此將可造福癌症病人。

關鍵字

抗口腔癌藥物 ； 太平洋紫杉醇 ； 艾黴素 ； 薑黃素 ； Memantine ； 人類口腔鱗狀細胞上皮癌細胞OECM-1 ； 藥物聯用指數 ； 效價比

並列摘要

The current incidence of oral cancer and mortality in Taiwan is becoming increasingly serious. According to the statistics of Ministry of Health and Welfare, oral cancer is the fifth of the top ten cancer deaths in 2015. The male mortality rate of oral cancer in Taiwan is 11.1 times that of women and the median age of death was 58 years, which is about 10 years younger than the median age of all cancer deaths.
Recent studies showed that the important factors involved in cancer cell formation are inflammation, oxidative stress and free radicals and activation of NMDA receptors, etc. Oral cancer is due to long-term oral stimulation by carcinogens or betel nut chewing which induce complicated carcinogenic processes. It has been estimated that about 90% of oral cancer patients in Taiwan have the habit of chewing betel nut.
Treatments of oral cancer include surgical resection, radiotherapy and chemotherapy. The commonly used chemotherapeutic agents such as 5-fluoro-uracil, cisplatin, paclitaxel (taxol), docetaxel (taxotere), gemcitabine (gemzar), vinorelbine (navelbine), capecitabine (xeloda) and so on. Because of the side effects (nausea, vomiting, myelosuppression decreased immunity) and drug resistance of these drugs, it is urgent to develop newer chemotherapeutic agents with better efficacies and less side effects.
Therefore, this study aimed at searching for drug combinations which composite different drugs with different action mechanisms. The drugs we chose for this study include taxol, curcumin, memantine and doxorubin. Taxol and doxorubin are clinical chemotherapeutic agents, curcumin is well known a phytopolyphenol with antioxidant, anti-inflammatory, anti-cancer and neuroprotective effects and memantine is an uncompetitive antagonist of NMDA receptor.
The effects of these drug combinations on the growth of oral squamous cell carcinoma cell line (OECM-1) and Smulow-Glickman gingival cells (SG) were studied by MTT assay. Combination index (CI) and potency ratio (P.R.) were calculated in order to analyze whether their combined effects are potentiated, additive or antagonistic. We found that inhibitory effect of anticancer drugs, taxol and doxorubicin, inhibition the growth of OECM-1 for longerterm is better than that of short-term effect. TC combination and TCM combination for the inhibition of OECM-1 proliferation are better than single-use and SG. When Taxol+Curcumin combination use 21hours, the CI=0.73, P.R.=1.4. When Taxol+Curcumin+Memantine combination use 43hours, the CI=0.64. They are synergistic. These drug combinations potentiate with each other in the inhibitory effects on OECM-1 proliferation. In the future, it is expected to have clinical trials to prove found in this study that the novel drug combinations have better anti-cancer effect and less side effects. So it will benefit cancer patients.

並列關鍵字

Anti-oral cancer drugs ； Taxol ； Doxorubicin ； Curcumin ； Memantine ； OECM-1 ； combination index ； potency ratio

參考文獻

2. Saman, W. (2009) Global epidemiology of oral and oropharyngeal cancer. Oral Oncology,45,309-16

3. Kevin D. S., Jacques F., Ahmedin J., Rengaswamy S., Chaturvedi., Freddie B.,& Isabelle S.(2017).The Global Incidence of Lip, Oral Cavity, and Pharyngeal Cancers by Subsite in 2012. CA CANCER J CLIN,67,51-64

4. Soria J.C., Kim E.S., Fayette J., Lantuejoul S., Deutsch E., & Hong W.K.(2003). Chemopreventive and Anti-cancer Efficacy of Silibinin against Growth and Progression of Lung Cancer. Lancet Oncol,4, 659-69.

5. Coussens L.M., & Werb Z. (2002).Nature,420, 860-7.

7. Stepulak A., Rola R., Polberg K., &Ikonomidou C.(2014).Glutamate and its Receptors in Cancer. J Neural Transm, 121(8),933-44.