| 摘要: | 近年來，由於高齡化及生活型態的改變，使得國人癌症發生人數仍持續增加。根據衛生福利部 (2016) 統計，台灣地區自 1982 年以來，癌症高居國人十大死因的首位，其中大腸直腸癌為癌症死亡原因第三大主因。牛樟芝 (Antrodia cinnanomea) 為台灣特有的藥用真菌，過去許多研究指出牛樟芝不僅具有保肝的功能且具有抗癌功效，因成分中富含三?類及多醣體，可抑制癌細胞生長或凋亡。近十年來，奈米科技用於保健食品的應用越來越廣泛，研究指出將食品奈米化後可增加該成分的吸收與利用，且中草藥奈米化及藥物奈米化已被證實可增加生物利用率及溶解度並且能減少藥物毒性、強化藥理活性等功能，因此本實驗欲比較一般研磨及奈米研磨牛樟芝子實體對於細胞凋亡作用之差異。本實驗以 MTT assay 比較一般研磨牛樟芝子實體 (Ground Antrodia cinnamomea fruiting body, GACFB) 及奈米研磨牛樟芝子實體 (Nanonized Antrodia cinnamomea fruiting body, NACFB) 對於四株癌細胞，大腸直腸癌 ( human colon adenocarcinoma cells, 如Colo205 and HT-29)、肺癌細胞 (human lung adenocarcinoma, A549)、肝癌細胞(liver hepatocellular cells, HepG2) 的毒殺作用，結果顯示 NACFB 對 HT-29 的毒殺作用具專一性且其 IC50 顯著低於 GACFB，以流式細胞儀分析癌細胞週期發現在 200 μg/mL 濃度下，NACFB 較 GACFB 更能增加 HT-29 細胞在 subG1、G1 與 S phase 的比例 (8.8、11.8 及 25.5%)，並降低 G2/M phase 18.3%。以濃度 200 g/mL 的 NACFB 或 GACFB 處理HT-29 細胞 24小時，在腫瘤抑制蛋白 p53 及促進細胞凋亡蛋白質 (Bcl-2 Assaciated X protein, Bax) 兩組無顯著差異；蛋白質表現量在 200 μg/mL 的濃度下 NACFB 組較 GACFB 組顯著降低在抗細胞凋亡蛋白質 (B cell lymphoma 2) 中；顯著增加在 caspase-3 (cysteinyl aspartate-spesific protease-3) 及 caspase-9 (cysteinyl aspartate-spesific protease-9) 蛋白質。綜合以上， NACFB 較 GACFB 對 HT-29 細胞具有更佳的抑制分裂及促凋亡功效。
Colorectal cancer (CRC) is the third leading cause of cancer deaths in Taiwan as per the 2011 statistics and ranks third in cancer-related mortality in the world. Antrodia cinnamomea, a Taiwan-specific medicinal mushroom, has been reported that possessed many biological activities, including hepatoprotection, antiinflammation, antihepatitis B virus and anticancer. Recently,the development of nanoherbal drugs possessing high bioavaibility and pharmacological activity , which consequently will open the new research area of the nanomization of herbal drugs against cancer and various other diseases. In the present study, ground Antrodia cinnamomea fruiting body (GACFB) and nanonized Antrodia cinnamomea fruiting body (NACFB) were used to compare apotosis effects on human colorectal adenocarcinoma cell line, HT-29 cells. The results from MTT assay indicated the IC50 of HT-29 cells treated by NACFB decreased 39.7% when compared with GACFB. The ratios of subG1, G1 and S phase of HT-29 cells after treating with 200 g/mL NACFB for 24 hr significantly increased by 8.8, 11.8, 25.5% respectively when comparied with GACFB at the same concentration. In NACFB treated HT-29 cells, the amounts of Bcl2 was significantly decreased by 43.5%, the levels of caspase-3 and caspase-9 were significantly increased by 20.88% and 21.56% when compared with GACFB treated HT-29 cells. The results from this study showed that ACFB has a better anti-proliferation and apoptotic activity on HT-29 cells after nanoization. |
