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 a...
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 $0.1-1.0{\mu}g/ml$, $0.001-0.01{\mu}g/ml$ and $0.01-0.1{\mu}g/ml$, 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 $0.01-1.0{\mu}g/ml$ 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.
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 $0.1-1.0{\mu}g/ml$, $0.001-0.01{\mu}g/ml$ and $0.01-0.1{\mu}g/ml$, 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 $0.01-1.0{\mu}g/ml$ 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.
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문제 정의
05). This is the first report on the antitumor effect of taxol produced from taxol-producing fungi on cancer cells, and our study offers important information and a new resource of producing this important anticancer drug by endofungus fermentation.
Therefore, more work is needed for a better understanding of the fungal taxol-induced cell apoptosis, such as the molecular mechanism of fungal taxol-induced cell apoptosis. This study offers important information and a new resource for the production of an important anticancer drug by endofungus fermentation to meet the increasing demand for taxol on the market.
제안 방법
, 2014). In this study, we investigated the effects of the fungal taxol produced from the strain HDFS4-26 on the proliferation and apoptosis of human breast cancer MCF-7 cells, cervical cancer HeLa cells and ovarian cancer HO8910 cells. We observed that the fungal taxol exhibited the ability of inducing growth inhibition and cell apoptosis in these tumor cells in a dose- and time-dependent manner.
To determine whether the fungal taxol can induce apoptosis, we treated HeLa, MCF-7 and HO8910 cells with 0.1 μg/ml or 0.01 μg/ml of fungal taxol for 48 h, and assessed the changes of cell apoptotic morphology, nuclear condensation and nuclear fragmentation by Geimsa stain and Hoechst 33258 staining, respectively.
대상 데이터
Cervical cancer HeLa cells were obtained from State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), the Chinese Academy of Agricultural Sciences (CAAS). Human breast cancer cell line MCF-7 and ovarian cancer cell line HO8910 were obtained from the Fourth Affiliated Hospital of Harbin Medical University, China.
, 2008). N. sylviforme HDFS4-26 (CCTCC M 208026) was deposited in the China Center for Type Culture Collection (Beijing, China).
Cell morphological change was observed with an EVOS digital inverted microscope (Bothell, WA, USA). Photographs were taken with a Nikon FM 10 camera.
데이터처리
Student’s t-test was used to compare the mean differences between samples by the statistical software SPSS version 10.0.
이론/모형
The fungal taxol was extracted from fermented fungal culture of stain HDFS4-26 by methanol and ethyl acetate method. The procedure of fungal taxol extraction, purification and analysis was carried out as previously described (Zhao et al.
성능/효과
05). The findings indicated that fungal taxol and yews-derived taxol have the same inhibitory effect on cancer cells. Meanwhile, fungal taxol significantly reduced cell viability of HeLa, MCF-7 and HO8910 cells (p<0.
The results showed that methanol at higher than 0.096 % for 24h or 48h affected significantly the proliferation and survival of HO8910 cells (p<0.05), methanol at higher than 0.0096 % for 72h affected significantly the proliferation and survival of HO8910 cells (p<0.05) (Table 1); and that methanol at higher than 0.0096 % for 24h or 48h affected significantly the proliferation and survival of MCF-7 and HeLa cells (p<0.05), methanol at higher than 0.00096 % for 72h affected significantly the proliferation and survival of MCF-7 cells (p<0.05) (Table 1).
Figure 3b, c and d showed similar results when analyzed by Hoechst33258 staining after treatment 48h by 10-flod increased fungal taxol. The results suggested that fungal taxol induced the concentration-and timedependent apoptosis of HeLa, MCF-7 and also HO8910 cells.
후속연구
Although our results in this study and previous reports have determined the effects of inducing apoptosis and the accordance of taxol from endophytic fungi and yew tree, there are many unclear aspects. Therefore, more work is needed for a better understanding of the fungal taxol-induced cell apoptosis, such as the molecular mechanism of fungal taxol-induced cell apoptosis. This study offers important information and a new resource for the production of an important anticancer drug by endofungus fermentation to meet the increasing demand for taxol on the market.
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