Cytotoxic effect of oyster mushroom Pleurotus ostreatus on human androgen-independent prostate cancer PC-3 cells
Cytotoxic effect of oyster mushroom Pleurotus ostreatus on human androgen-independent prostate cancer PC-3 cells
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DOI：
10.1089/jmf.2006.9.196
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发表时间：
2006-06-01
影响因子：
2.4
通讯作者：
Sivam, Gowsala
中科院分区：
文献类型：
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作者：
Gu, Yti-Huan;Sivam, Gowsala
本研究筛选了20种食用菌和3种纯化的食用菌多糖对人雄激素非依赖性肿瘤PC-3细胞的抗肿瘤活性。在所测试的蘑菇物种中，从新鲜平菇中制备的水溶性提取物（POE）对PC-3细胞产生最显着的细胞毒性。同时，用Annexin V-荧光素异硫氰酸酯流式细胞仪检测，POE（150 μ g/mL）作用PC-3细胞2 h后，可诱导PC-3细胞迅速凋亡。诱导的细胞凋亡也通过DNA片段末端脱氧核苷酸转移酶介导的X-dUTP缺口末端标记染色来证实，同时将POE（200 μ g/mL）加入PC-3细胞中6小时。POE对PC-3细胞的毒性和诱导凋亡均呈剂量依赖性。有趣的是，PC-3细胞在非贴壁生长条件下似乎对POE更敏感。与不含POE的培养基（定义为100%）相比，在添加POE（60或120 μ g/mL）的软琼脂培养基中肿瘤集落形成效率显著降低至4.5%或0.5%。POE加工过程中的温度对细胞毒活性起着决定性作用。在高温（80摄氏度）下暴露2小时，POE的生物活性就会消失;但在40摄氏度以下的一系列温度下，它仍然保持稳定。通过高效液相色谱的尺寸排阻和CellTiter 96（R）AQueous细胞增殖测定（Promega，麦迪逊，WI）分析和鉴定活性级分POE-F2。由于POE-F2也对热敏感并且具有强的280 nm吸收，因此结果暗示从糙皮侧耳中回收的活性化合物是水溶性蛋白质或多肽。
Twenty species of edible mushrooms and three purified mushroom polysaccharides were screened for their antitumor potential on human androgen-independent cancer PC-3 cells. A water-soluble extract (POE) prepared from the fresh oyster mushroom Pleurotus ostreatus produced the most significant cytotoxicity on PC-3 cells among the mushroom species tested. At the same time, POE induced a rapid apoptosis on PC-3 cells detected with annexin V-fluorescein isothiocyanate flow cytometry when the cells were exposed to POE (150 mu g/mL) for 2 hours. Induced apoptosis was also confirmed by DNA fragment terminal deoxynucleotidyl transferase-mediated X-dUTP nick end labeling staining while POE (200 mu g/mL) was added to PC-3 cells for 6 hours. Both cytotoxicity and induced apoptosis mediated by POE in PC-3 cells are dose-dependent. Interestingly, PC-3 cells appeared to be more sensitive to POE in anchorage-independent growth condition. Tumor colony-forming efficiency was dramatically reduced to 4.5% or 0.5% in POE (60 or 120 mu g/mL)-supplemented soft agar medium compared with that of POE-free medium (defined as 100%). Temperature in POE processing plays a decisive role for the cytotoxic activity. Bioactivity of POE was eliminated by exposure to high temperature (80 degrees C) for 2 hours; however, it remained stable at a series temperatures of below 40 degrees C. The active fraction POE-F2 was analyzed and identified by size exclusion of high performance liquid chromatography and the CellTiter 96 (R) AQueous Cell Proliferation Assay (Promega, Madison, WI). Since POE-F2 is also sensitive to heat and has strong 280 nm absorption, the results imply that active compounds recovered from P. ostreatus are water-soluble proteins or polypeptides.