陳明陽鄭淑蓉陳啟楨陳健祺2014-10-272014-10-272005-06-??http://rportal.lib.ntnu.edu.tw/handle/20.500.12235/6777硫磺菌(Laetiporus sulphureus)在傳統的中醫治療中可作為乳腺癌、前列腺癌之輔助食物,它可以幫助病患對抗癌症,但其作用機制尚未釐清?本實驗探討在不同培養條件下對硫磺菌生長速率之影響,在實驗過程中利用固態培養基探討最適培養硫磺菌之pH值、溫度及培養基,再以液態培養探討最適生長培養基之碳氮源組成及比例。結果顯示,在24℃、pH6.0下以0.5%蛋白腖(peptone)加3%麥牙糖抽出物(malt extract)為培養基,菌絲體生長狀況最好。利用液態培養所得之硫磺菌發酵液處理腫瘤細胞,結果顯示,隨著發酵天數遞增,對抑制腫瘤細胞的增生能力有顯著遞增現象,第十二天硫磺菌發酵液抑制腫瘤細胞(HeLa、Hep G2、HT1080、DU-145、MCF-7以及MDA-MB-231)的增生能力皆大於75%。硫磺菌發酵液處理HT1080細胞後以gelatin zymography analysis分析,結果發現硫磺菌發酵液具有顯著地抑制HT-1080細胞分泌金屬蛋白酶-9(metalloproteinase-9)的能力。因此本實驗的結果顯示,硫磺菌之液態發酵液不僅具有腫瘤細胞的增生,同時也具有抑制腫瘤細胞轉移的功能,由此可知,硫磺菌是值得進一步探討的食用菇類。Laetiporus sulphureus has been used in Chinese traditional medicine for a long time. However, there are very few scientific evidence to support its biological functions. Therefore, the optimal culture conditions for L. sulphureus, such as pH or temperature, were investigated in this study. The biological activities of cultural broth and mycelium of L. sulphureus, including antitumor activity awl antimetastasis, ere evaluated. The growth inhibition in tumor cells was evaluated by MTT-microculture tetrazolium assay and the ability of anti-metastasis was investigated by Gelatin Zymography analysis. The results showed that the optimal temperature was under 24℃ and optimal pH was around 6, and the best nutrient composition was 0.5% peptone +3% malt extract. Moreover, the liquid culture of L. sulphureus at day 12 showed the best inhibitory effect on the growth of tumor cells, such as HeLa. Hep G2, HT1080, DU-145, MCF-7 and MDA-MB-231 cells, by almost more than 75%. Furthermore, Gelatin Zymography analysis revealed that the expression of metalloproteinase-9 (MMP-9) in HT1080 cells treated with day-12 cultured broth of L. sulphureus was significantly suppressed. This result reveals that it is worthy to do more studies to illustrate the bioactivities of L. sulphureus.硫磺菌菌絲體液態培養腫瘤細胞抑癌試驗金屬蛋白酶-2金屬蛋白酶-9轉移Laetiporus sulphureusMyceliumLiquid cultureSolid cultureAntitumorMTTGelatin zymographyMetalloproteinaseMMP-9AntimetastasisThe Study of Liquid Culture and Bioactivity in Laetiporus Sulphureus硫磺菌(Laetiporus Sulphureus)之液態培養及其生理活性研究