Wang Xin, Wang Chao, Sun Yu-Ting, Sun Chuan-Zhen, Zhang Yue, Wang Xiao-Hua, Zhao Kai
Key Laboratory of Microbiology, School of Life Science, Heilongjiang University, Harbin, China E-mail :
Asian Pac J Cancer Prev. 2015;16(1):125-31. doi: 10.7314/apjcp.2015.16.1.125.
Currently, taxol is mainly extracted from the bark of yews; however, this method can not meet its increasing demand on the market because yews grow very slowly and are a rare and endangered species belonging to first- level conservation plants. Recently, increasing efforts have been made to develop alternative means of taxol production; microbe fermentation would be a very promising method to increase the production scale of taxol. To determine the activities of the taxol extracted from endophytic fungus N. sylviforme HDFS4-26 in inhibiting the growth and causing the apoptosis of cancer cells, on comparison with the taxol extracted from the bark of yew, we used cellular morphology, cell counting kit (CCK-8) assay, staining (HO33258/PI and Giemsa), DNA agarose gel electrophoresis and flow cytometry (FCM) analyses to determine the apoptosis status of breast cancer MCF-7 cells, cervical cancer HeLa cells and ovarian cancer HO8910 cells. Our results showed that the fungal taxol inhibited the growth of MCF-7, HeLa and HO8910 cells in a dose-and time-dependent manner. IC50 values of fungal taxol for HeLa, MCF-7 and HO8910 cells were 0.1-1.0 μg/ml, 0.001-0.01 μg/ml and 0.01- 0.1 μg/ml, respectively. The fungal taxol induced these tumor cells to undergo apoptosis with typical apoptotic characteristics, including morphological changes for chromatin condensation, chromatin crescent formation, nucleus fragmentation, apoptotic body formation and G2/M cell cycle arrest. The fungal taxol at the 0.01-1.0 μg/ ml had significant effects of inducing apoptosis between 24-48 h, which was the same as that of taxol extracted from yews. This study offers important information and a new resource for the production of an important anticancer drug by endofungus fermentation.
目前，紫杉醇主要从红豆杉树皮中提取；然而，由于红豆杉生长非常缓慢，且属于国家一级保护植物中的珍稀濒危物种，这种方法无法满足市场对其日益增长的需求。最近，人们加大了开发紫杉醇生产替代方法的力度；微生物发酵将是一种非常有前景的提高紫杉醇生产规模的方法。为了确定从内生真菌N. sylviforme HDFS4 - 26中提取的紫杉醇在抑制癌细胞生长和诱导癌细胞凋亡方面的活性，与从红豆杉树皮中提取的紫杉醇进行比较，我们采用细胞形态学、细胞计数试剂盒（CCK - 8）检测、染色（HO33258/PI和吉姆萨染色）、DNA琼脂糖凝胶电泳和流式细胞术（FCM）分析来确定乳腺癌MCF - 7细胞、宫颈癌HeLa细胞和卵巢癌HO8910细胞的凋亡状态。我们的结果表明，真菌紫杉醇以剂量和时间依赖性方式抑制MCF - 7、HeLa和HO8910细胞的生长。真菌紫杉醇对HeLa、MCF - 7和HO8910细胞的IC50值分别为0.1 - 1.0μg/ml、0.001 - 0.01μg/ml和0.01 - 0.1μg/ml。真菌紫杉醇诱导这些肿瘤细胞发生凋亡，具有典型的凋亡特征，包括染色质浓缩、染色质新月形成、细胞核碎片化、凋亡小体形成和G2/M细胞周期阻滞。0.01 - 1.0μg/ml的真菌紫杉醇在24 - 48小时之间具有显著的诱导凋亡作用，这与从红豆杉中提取的紫杉醇相同。本研究为通过内生真菌发酵生产重要抗癌药物提供了重要信息和新资源。
