Effect Of Sulforaphane On Autophagy In A375 Cells Of Melanoma

ObjectiveTo investigate the effect of SFN on the autophagy of HMM A375 cells and the function of autophagy in inhibiting proliferation of SFN tumor cells.MethodsThe proliferation rate（PI）of human melanoma A375 cells was measured by MTT method after being treated with SFN of different concentrations(0M,5×10^-6M,1×10^-5M,2×10^-5M,4×10^-5M)for one day.The autophagy was determined by real-time PCR as LC3,beclin-1,p62 Western blotting was used to detect the expression of LC3,beclin-1 and p62.Chloroquine（CQ）,a 20μmol·L-1 autophagy inhibitor,was used to pretreat human melanoma A375cells for 120 min,and then SFN(0 M,5×10^-6 M,1×10^-5M,2×10^-5 M,4×10^-5 M)of various concentrations was used to treat human melanoma A375 cells for one day,and the cell proliferation rate was measured by MTT method.ResultsFor human melanoma A375 cell proliferation activity,SFN could play a significant inhibitory effect.At the same time,with the increase of SFN treatment dose,A375 cell activity was significantly weakened,showing a dose-response relationship（P<0.05,P<0.01）.Compared with the blank group,as the SFN intervention dose increased,the ratio of LC3 m RNA and LC3Ⅱ/LC3Ⅰprotein expression and Beclin-1 m RNA and protein expression also increased,but P62 m RNA and protein expression decreased（P<0.05）.After A375 cells were treated with autophagy inhibitor CQ,the cell proliferation activity of the SFN+CQ group was significantly lower than that of the SFN alone treatment group（P<0.05）.ConclusionsSFN can inhibit the proliferation of A375 cells,and can also promote autophagy,while inhibiting autophagy can increase the inhibitory effect of SFN on the proliferation of A375 cells of melanoma.