2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38044Cancer is now a major worldwide disease, accounted for over 7 million deaths per year. The majority (60-90%) of human cancer is considered to be attributed to external factors such as environment, diet, infection and other life style factors. Only few cases might result from inherent genetic alteration. Thereafter it is important to understand the toxicological origins and bio-pharmacology of environments. It is reported that carcinogenesis is a multiple step process characterized into 3 stages including initiation, promotion, and progression. Promotion is an operational event that leads to sustained preneoplastic lesion and subsequently development to neoplasm. Oxidative stress and inflammation are supposed to be a critical step related to tumor promotion. Since both arsenic and humic acid induces oxidative stress we will evaluate their tumor promotion effect and association in this study. Inorganic arsenics such as arsenite and arsenate have been shown epidemiologically to be carcinogenic; nevertheless, experimental animal studies have not conclusively proven their carcinogenicity. Recently, it is reported that arsenite induced cell transformation and dimethylarsinic acid (DMA), a main metabolite of inorganic arsenics, to promote tumorigenesis in an experimental model using rodents. Humic acid is a group of polymers with high molecular weights that are the results of decomposition of organic matter, especially dead plants. It was implicated as a causal factor of cancer, goiter, Blackfoot disease and Kashin-Beck disease. Since epidemiological studies reported an explicit association between arsenic and humic acid in ground water and dermatological diseases and skin cancer. We propose to set up evaluation method of humic acid free radicals and/or heavy metal by chemiluminescence system, then use two-step skin carcinogenesis model to study the association of humic acid and arsenic as a tumor promoter. The following topics will fulfill: The first year: 1. Isolation and analysis of humic acid (HA-N) from soil of Central Mountain in Taiwan. 2. Synthesis and analysis humic acid (HA-S). 3. Set up evaluation method of humic acid free radicals and/or toxic substance from environment such as heavy metal by chemiluminescence system. 4. Transformation evaluation of arsenite or DMA in JB6 cells. 5. Determination Cell cycle distribution of arsenite or DMA in JB6 cells. The second year: 1. Evaluation transformation potential of arsenic with humic acid (HA-N or HA-S) in JB6 cells. 2. Evaluation cell cycle distribution of arsenic with humic acid (HA-N or HA-S) in JB6 cells. 3. Evaluation transformation potential and cell cycle distribution of arsenic with humic acid from groundwater (from other co-plan) in JB6 cells. 4. Evaluation proliferation signal of arsenic with humic acid in JB6 cells. 5. Evaluation AP-1 trans-activation of arsenic with humic acid in JB6 cells. The third year: 1. Effect of arsenite and DMA induced ornithine decarboxylase and myeloperoxidase activation in CD-1 mice skin. 2. Effect of arsenite and DMA induced ear edema and skin inflammation in CD-1 mice skin. 3. Effect of humic acid (HA-N or HA-S) on arsenic (arsenite or DMA) induced ornithine decarboxylase and myeloperoxidase activation in mice skin. 4. Effect of humic acid (HA-N or HA-S) on arsenic induced ear edema and skin inflammation in CD-1 mice skin. 5. Effect of humic acid (HA-N or HA-S) on arsenic induced two-step skin tumor model. (bezo(a)pyrene as a initiatior; arsenic as a promoter).癌症是目前世界各國普遍主要死亡病因之一，每年大約有七百萬人死於癌 症，而根據研究又認為百分之六十至九十人類之癌症是因外在因素如環境、飲食 等所引起，因此了解環境之變化與危險因子，進而加以避免、預防成為控制癌症 發生之重要課題。砷是地球表面無處不存在的元素，由於岩石和土壤中礦物質的 日益風化消蝕，因此使得無機砷被釋放進入地下水層，造成砷的污染，飲用過量 砷的地下水和地方性疾病例如烏腳病、皮膚癌等的發生有密切的關係，雖然流行 病學顯示砷與很多癌症之發生有關，体外實驗也證實其為一致癌物，但至今仍無 一成功之動物致癌模式，因此砷在癌化過程之角色有待釐清; 腐植酸是環境中一 種酚類聚合物，主要來自於死亡植物之分解產物，富含於有機肥料、土壤、地下 水中，台灣及中國大陸學者近40 年來研究證實其與克山病、大骨節病、甲狀腺 腫及烏腳病等地方性疾病有關，另有研究顯示烏腳病流行地區之癌症累積致死率 也相當高，一些體內或體外研究顯示腐植酸為無(或弱)基因毒性，因此腐植酸在 癌化過程角色有待進一步探討。 近年來科學家對於癌化之過程有了進一步之了解，發現癌化過程需經多重步 驟，其中包括: 起始作用(initiation), 促進作用(promotion), 擴展作用(progression and expansion)等階段，又很多報告指出活性氧系具有癌促進作用，最近文獻指出 腐植酸、無機砷及其代謝產物dimetthylarsinic acid (DMA)可產生氧化壓力損傷或 氧化性毒性，另低劑量無機砷可促進JB6 細胞之轉型，口服DMH 可促進dimethyl benz(a)anthracene (DMBA) 誘發皮膚腫瘤之形成，因此本研究擬先由萃取天然或 合成之腐植酸以化學微量發光儀建立腐植酸與環境中有害物質產生自由基交互 作用之評估方法，再比較無機砷及其代謝產物dimethylarsinic acid (DMA)之促進 JB6 細胞轉型及促進dimethylbenz(a) - anthracene (DMBA) 誘發皮膚腫瘤形成作 用之差異，進而探討腐植酸對砷之此二活性之影響。研究項目如下: 第一年 1. 由中部過度開發高山之深層土壤萃取腐植酸(HA-N)並以自由基測定儀等儀 器測定分析。 2. 利用protocatechuic acid 合成腐植酸(HA-S)並以自由基測定儀等儀器測定 分析。 3. 以化學微量發光儀建立腐植酸對環境中有害物質如重金屬等產生自由基交 互作用之評估。 4. 比較無機砷及其代謝產物DMA 對細胞週期之影響。 5. 比較無機砷及其代謝產物dimethylarsinic acid (DMA)促進JB6 細胞轉型能 力之異同。 第二年 1. 比較有無腐植酸(HA-N or HA-S)對砷促進JB6 細胞週期變化之影響。 2. 比較有無腐植酸(HA-N or HA-S)對砷促進JB6 細胞轉型能力之影響。 3. 比較其它子計畫所提供地下水腐植酸對砷促進JB6 細胞週期變化之影響。 4. 比較其它子計畫所提供地下水腐植酸對砷促進JB6 細胞轉型能力之影響。 5. 由增生訊息路徑及轉譯因子之活性探討腐植酸對砷促進JB6 細胞轉型影響 之分子機制。 6. 將HA-N 及HA-S 送其它子計劃進行分離,依transformation assay 評估出 作用最強之化合物,定出結構並送其它子計畫進行其它毒理之研究。 第三年 1. 比較無機砷及其代謝產物dimethylarsinic acid (DMA) 活化老鼠皮膚 ornithine decarboxylase (與polyamine metabolism 有關)及myeloperoxidase 作用之異同。 2. 比較無機砷及其代謝產物dimethylarsinic acid (DMA) 誘發皮膚發炎及耳朵 水腫作用之異同。 3. 比較腐植酸(HA-N or HA-S) 對有無砷活化下老鼠皮膚ornithine decarboxylase (與polyamine metabolism 有關)及myeloperoxidase 作用之差 異。 4. 比較腐植酸(HA-N or HA-S)對有無砷誘發下皮膚發炎及耳朵水腫作用之差 異。 5. 比較有無腐植酸(HA-N or HA-S)對Benzo(a)pyrene 為起始物砷為促進物誘 發皮膚腫瘤作用之影響。 6. 對其它子計劃分離出有意義之化合物以本研究模式進行測試。砷腐植酸腫瘤促進作用Arsenichumic acidtumor promotionProject