Investigating Migration Inhibition and Apoptotic Effects of Fomitopsis pinicola Chloroform Extract on Human Colorectal Cancer SW-480 Cells

Abstract

Background Fomitopsis pinicola (Sw. Ex Fr.m) Karst (FPK) which belongs to the Basidiomycota fungal class is one of the most popular medical fungi in China. It has been used for many diseases: cancer, heart diseases, diabetes and so on. However, little study on the pro-apoptotic effect and migration inhibition of FPK chloroform extract (FPKc) has been reported and the possible involved mechanism has not been illuminated.Methodology/Principal FindingsChemical analysis was performed by HPLC which showed ergosterol (ES) concentration was 105 µg/mg. MTT assay revealed that FPKc could selectively inhibit SW-480 cells viability with the IC50 of 190.28 µg/ml. Wound healing and transwell assay indicated that 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. 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 up-regulate P53 expression and thus lead to G1 phase arrest. When SW-480 cells were pretreated with N-acetylcysteine (NAC), the ROS generation, cell viability and apoptotic ratio were partially declined, which indicated that ROS was vertical in the pro-apoptosis process induced by FPKc. Moreover, in the whole process, ES which has been previously found in FPKc had the similar effect to FPKc. Thus we could conclude that ES, as one of the highest abundant components in FPKc, might also be one of the active constituents.Conclusion/SignificanceFPKc could inhibit the migration of SW-480 cells, induce SW-480 cells G1 phase arrest and cause ROS-mediated apoptosis effect. And ES might be one of the effective constituents in the whole process.

The Antitumor Activity of Meconopsis Horridula Hook, a Traditional Tibetan Medical Plant, in Murine Leukemia L1210 Cells

“…The effect of M. horridula extract on L1210 cell viability was determined with the conventional MTT (Sigma, St Louis, USA) reduction assay [18]. Briefly, 100 µL L1210 cell suspension was placed in a 96-well culture plate at a density of 1×10 5 cells/mL and incubated with various concentrations of M. horridula (0, 30, 60, 90 and 120 µg/mL) extract for distinct time points (24 h and 48 h).…”

“…The intracellular ROS level was evaluated by the oxidative conversion of 2′, 7′-dichlorofluoresceindiacetate (DCFH-DA) to fluorescent 2′, 7′-dichlorofluorescein (DCF) [18]. L1210 cells were treated with the mentioned concentrations of M. horridula extract for 2 and 6 h. The treated cells were harvested, washed twice with PBS, re-suspended in 500 µl of 10 µM DCFH-DA (Molecular Probes Inc., Invitrogen, CA, USA) and incubated at 37 ℃ for 30 min in the dark.…”

“…Many natural products have been reported to inhibit cancer cell proliferation by increasing intracellular ROS level. For instance, Fomitopsis pinicola chloroform extract induced SW-480 cell apoptosis by increasing ROS accumulation [18]. Acidic PAP-3 (a polysaccharide) has also been shown to exert anti-cancer effects through the ROS-mediated mitochondrial apoptotic pathway [19].…”

In Vitro and In Vivo Activity of Fomitopsis Pinicola (Sw. Ex Fr.) Karst Chloroform (Fpkc) Extract Against S180 Tumor Cells

“…If this type of arrest cannot be repaired, cells may undergo apoptosis, which is a programmed cell death characterized by DNA degradation, chromatin condensation, and nuclear fragmentation [24]. Deregulation of apoptosis is significant in the initiation and progression of many cancers so that apoptosis induction could be the most promising defense against cancer development [24-28]. In our study, we used HO-PI staining and Annexin V-FITC/PI assay to clarify the type of cell death induced by FPKc.…”

Chemical composition of the mushroom Meripilus giganteus Karst. and bioactive properties of its methanolic extract

“…Antimigratory potential of mushroom's extracts has been reported for other classes like Fomitopsis pinicola which belongs to the Basidiomycota fungal class that could inhibit the migration of colon cancer cell line SW-480 (Wang et al, 2014), or Ganoderma lucidum that exerts strong antimigratory effect on ovarian cancer cell lines (Zhao et al, 2011). This is the first report presenting antimigratory potential of M. giganteus extract on cancer cells in vitro.…”