关键词: 黑色素瘤, 莱菔硫烷, 自噬, 氯喹

Abstract:

Objective To investigate the effect of sulforaphane（SFN） on autophagy of human melanoma A375 cells. Methods The proliferation of A375 cells was detected by MTT assay after treated with SFN at different concentrations（0 μmol·L^-1 ，5 μmol·L^-1，10 μmol·L^-1，20 μmol·L^-1，40 μmol·L^-1） and its combined autophagy inhibitor chloroquine. Theexpressions of autophagy-related molecules LC3（LC3Ⅱ/LC3Ⅰ），Beclin-1 and p62，apoptosis-related molecules Bcl-2，Bax and Caspase-3 were detected by Real-time PCR and Western blot. Results MTT assay showed that SFN inhibited the proliferation of A375 cells in a dose-dependent manner（F=21.517，P<0.001）. Compared with the control group，SFN at different concentrations（5 μmol·L^-1，10 μmol·L^-1，20 μmol·L^-1，40 μmol·L^-1） could up-regulate the expression of autophagy-related molecules LC3Ⅱand Beclin-1 mRNA，LC3Ⅱ/LC3Ⅰand Beclin-1 proteins，as well as the expression of pro-apoptotic proteins Bax and Caspase-3 （P<0.05），while down-regulated the expression of autophagy-related molecule p62 and the inhibitor of apoptosis protein Bcl-2（P<0.05）. The proliferation rate of A375 cells treated with chloroquine combined with SFN at different concentrations was lower than that treated with SFN alone（P<0.05）. Conclusions SFN can induce autophagy and promote apoptosis of melanoma A375 cells，and can enhance the inhibition of cell proliferation when combined with chloroquine，and can be an effective therapeutic for melanoma.

Key words: Melanoma, Sulforaphane, Autophagy, Chloroquine