RESUMO
The purpose of this study was to investigate the effect of Huaier on autophagy of human hepatoma SK-HEP-1 cells and the effect of autophagy on the proliferation of SK-HEP-1 cells. CCK-8 assay was used to evaluate the effect of Huaier on the proliferation of SK-HEP-1 cells under different concentrations and different times. Acridine orange staining was used to measure the effect of Huaier on the autolysosome formation in SK-HEP-1 cells. Immunofluorescence assay was applied to examine the effect of Huaier on the expression and distribution of autophagy marker LC3 in SK-HEP-1 cells. In additionï¼ LC3 expression was also checked by immunoblot analysis in the presence of Huaier. At lastï¼ the effects of Huaier in combination with autophagy inhibitor bafilomycin A1 on the proliferation of SK-HEP-1 cells was detected by CCK-8 assay. The results showed that Huaier aqueous extract significantly inhibited the proliferation of human hepatoma SK-HEP-1 cells in a dose- and time-dependent manner. Huaier aqueous extract dramatically promoted the formation of autolysosome in SK-HEP-1 cells. Moreoverï¼ Huaier markedly increased the number and intensity of intracellular LC3 fluorescent puncta and up-regulated LC3-â ¡ expression. These data indicated that Huaier evidently activated autophagy of SK-HEP-1 cells. Additionallyï¼ autophagy inhibition significantly attenuated the sensitivity of SK-HEP-1 cells to Huaier treatment. Thereforeï¼ autophagy activation is involved in the inhibitory effects of Huaier on the proliferation of human hepatoma SK-HEP-1 cells.