| Bacgarouds and objectivesFomitopsis pinicola (Sw. Ex Fr.) Karst (FPK) is a brown-rot fungus and has been widely distributed in the word. As one of the most popular traditional medical fungi, FPK has been used in many countries, such as in Korea, Japan, China, and so on. In western countries, FPK has been used for the treatment of headache and liver problems. In China, it has been used as a haemostatic, anti-diabetic and anti-inflammate agent in folk medicine. What’s more, the anticancer effect of FPK ethonal extract has also been studied in recent years. Thus, this study manily focused on the chemical analysis and anticancer funtion of FPK chloroform extract (FPKc) in vitro and in vivo, hoping to apply the promising medical fungi to clinical.Methods and resultsFirstly, HPLC assay was employed in this study. Chemical analysis indicated FPKc might contain ES (ergosterol), PA (Pachymic acid) and DA (Dehydroeburiconic acid). The percentage of them was 10.5%,3.56% and 0.25%, respectively. What’s more, after FPKc treatment, ES, as one of the main component of FPKc, could enter into intracellular of SW-480 cells to perform its function.Secondly, MTT assay revealed FPKc could inhibit viability of SW-480, SW-620 and Caco-2 cells in a time- and dose-dependent manner. Different cancer cells performed different sensitivities to FPKc. After 48 h incubation of FPKc, IC50s of SW-480 and SW-620 cells were 190.28 and 143.26 μg/ml, respectively. Caco-2 cells performed much less sensitive to FPKc than the other two cell lines. Annexin V-FITC/PI staining, nuclear Hoechst 33342 staining and DNA fragmentation analysis revealed that FPKc and ES could induce SW-480 cells apoptosis. The apoptosis process closely involved in ROS accumulation and depletion of GSH, activation of caspase 3, poly (ADP-ribose) polymerase (PARP) degradation. FPKc could also lead to G1 phase arrest. When SW-480 cells were pretreated with N-acetylcysteine (NAC), ROS generation, cell viability and apoptotic ratio were partially declined, which indicated ROS was vertical in the pro-apoptosis process induced by FPKc. In the whole investigation, ES performed similar anticancer effect with FPKc, suggesting ES might be one of the active compunds in FPKc.Thirdly, results of wound healing and transwell assay showed FPKc could inhibit the migration of SW-480 cells obviously. FPKc could also dramatically decreased the matrix metalloproteinases-2,9 (MMP-2 and MMP-9) expression. Moreover, in the whole process, ES, which has been previously found in FPKc, also could partialy inhibit the migration of SW-480 cells.At last, the obtained data indicated FPKc could obviously inhibit the solid tumor growth and prolong the survival time of tumor-bearing mice. What’s more, FPKc displayed little damage on normal cells in vitro and even on normal tissues in vivo.ConclusionsThis study was the first trial to investigate the pro-apoptotic and anti-migration function of FPKc on SW-480 cells. HPLC analysis showed FPKc contained ES, PA and DA; ES could enter SW-480 cells to perform its anticancer function. Our study showed FPKc could induce SW-480 cells apoptosis, cell cycle arrest and migration inhibition. The underlying mechanism detection revealed ROS might be the key factor in FPKc-induced apoptosis. In vivo, we found FPKc could obviously inhibit the solid tumor growth and prolong the survival time of tumor-bearing mice without damaging the normal tissuses.
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