Background： The suillin isoform iso-suillin is a natural substance isolated from a petroleum ether extract of the fruiting bodies of the mushroom Suillus.flavus. Previous studies have found its inhibition effect on s...Background： The suillin isoform iso-suillin is a natural substance isolated from a petroleum ether extract of the fruiting bodies of the mushroom Suillus.flavus. Previous studies have found its inhibition effect on some cancer cells, and we aimed to study its effects on human small cell lung cancer H446 cell line. Methods： Cell viability was measured by 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay. Cellular morphological changes （apoptosis and necrosis） were evaluated using an electron microscope and Hoechst 33258 staining detected by the inverted microscope. Flow cytometry was used to detect cell apoptosis, cell cycle distribution, and mitochondrial membrane potential. Protein expression was determined by Western blotting analysis. Results： Here, we describe the ability of iso-suillin to inhibit the growth of H446 cells in time- and dose-dependent way. Iso-suillin had no obvious impact on normal human lymphocyte proliferation at low concentrations （9.09, 18.17, or 36.35 μmol/L） but promoted lymphocyte proliferation at a high concentration （72.70 μmol/L）. After treatment of different concentrations of iso-suillin （6.82, 13.63, or 20.45 μmol/L）, the apoptosis rate of H446 cells increased with increasing concentrations of iso-suillin （16.70%, 35.54%, and 49.20%, respectively, all P 〈 0.05 compared with the control）, and the expression of related apoptotic proteins in the mitochondrial pathway including cytochrome c and caspase-9 were up-regulated compared with the control （all P 〈 0.05）. On the contrary, Bcl-2/Bax ratio was down-regulated compared with the control. Besides, the expression of pro-apoptotic proteins in the death receptor apoptosis pathway, including Fas-associating protein with a novel death domain and caspase-8, and the expression of caspase-3, a downstream regulatory protein of apoptosis, were also increased compared with the control （all P 〈 0.05）. lnhibitors of caspase-9 and caspase-8 reversed the apoptosis process in H446 展开更多
基金Financial support and sponsorship This study was supported by a grant from the Natural Science Foundation of Hebei Province, China （No. H2015206214）.
文摘Background： The suillin isoform iso-suillin is a natural substance isolated from a petroleum ether extract of the fruiting bodies of the mushroom Suillus.flavus. Previous studies have found its inhibition effect on some cancer cells, and we aimed to study its effects on human small cell lung cancer H446 cell line. Methods： Cell viability was measured by 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay. Cellular morphological changes （apoptosis and necrosis） were evaluated using an electron microscope and Hoechst 33258 staining detected by the inverted microscope. Flow cytometry was used to detect cell apoptosis, cell cycle distribution, and mitochondrial membrane potential. Protein expression was determined by Western blotting analysis. Results： Here, we describe the ability of iso-suillin to inhibit the growth of H446 cells in time- and dose-dependent way. Iso-suillin had no obvious impact on normal human lymphocyte proliferation at low concentrations （9.09, 18.17, or 36.35 μmol/L） but promoted lymphocyte proliferation at a high concentration （72.70 μmol/L）. After treatment of different concentrations of iso-suillin （6.82, 13.63, or 20.45 μmol/L）, the apoptosis rate of H446 cells increased with increasing concentrations of iso-suillin （16.70%, 35.54%, and 49.20%, respectively, all P 〈 0.05 compared with the control）, and the expression of related apoptotic proteins in the mitochondrial pathway including cytochrome c and caspase-9 were up-regulated compared with the control （all P 〈 0.05）. On the contrary, Bcl-2/Bax ratio was down-regulated compared with the control. Besides, the expression of pro-apoptotic proteins in the death receptor apoptosis pathway, including Fas-associating protein with a novel death domain and caspase-8, and the expression of caspase-3, a downstream regulatory protein of apoptosis, were also increased compared with the control （all P 〈 0.05）. lnhibitors of caspase-9 and caspase-8 reversed the apoptosis process in H446
