Effect Of Beclin1 Overexpression On Invasion And Migration Of Melanoma Cell

Purpose:To construct a melanoma cell line overexpressing Beclin1 and observe the changes of invasion and migration ability of SK-MEL-2 cells after overexpressing Beclin1.Methods:The expression of Beclin1 in pigmented nevus and melanoma tissues was observed by immunohistochemical SP method.The expression levels of Beclin1 in melanoma cells A375 and SK-MEL-2 were detected by Western blotting,and SK-MEL-2 cell lines with low expression of Beclin1 protein were screened as research objects.SK-MEL-2 cells were transfected with liposome and the transfection effect was identified by Western blotting.The changes of invasion and migration ability of SK-MEL-2 cells before and after transfection were detected by Transwell chamber method and cell wound healing method.Results:Beclin1 was expressed in pigmented nevus and CMM.The positive staining of Beclin1 was brown and dispersed in the cytoplasm and nucleus.The positive rate of Beclin1 protein expression in CMM was 37.14%,which was lower than that in pigmented nevus(72.00%).The difference was statistically significant(P < 0.05);The relative expression of Beclin1 protein in SK-MEL-2 cells was 0.04±0.02,which was significantly lower than that in A375 cells(0.67±0.15),with statistical significance(P<0.05);The expression of Beclin1 in pcDNA 3.1-Beclin1 group was 0.32 ±0.04,significantly higher than that in blank group(0.07 ±0.02)and pcDNA 3.1 group(0.06±0.02),with significant difference(P<0.05).There was no significant difference between untreated group and pcDNA 3.1 group(P>0.05).4.In pcDNA3.1-Beclin1 group,the number of cells passing through the cell chamber was 18.67±1.19,which was significantly lower than that in blank group(86.78±5.93)and pcDNA3.1 group(87.89±6.05).The difference was statistically significant(P<0.05).5.The 24-hour migration distance of pcDNA 3.1-Beclin1 group was 153.00±21.63 um,which was significantly lower than that of blank group(365.67±16.86 um)and pcDNA 3.1 group(367.67±47.69um)(P<0.05).The 48-hour migration distance of pcDNA 3.1-Beclin1 group was 340.33±10.60,which was significantly lower than that of blank group(340.33±10.60 um)and pcDNA 3.1 group(588.00±28.16um).The difference was statistically significantg(P < 0.05).Conclusions : Beclin1 protein was significantly down-regulated in cutaneous melanoma;Beclin1 protein was significantly low expressed in SK-MEL-2 cell line.SK-MEL-2 cell line with low expression of Beclin1 was selected for further study;The expression of Beclin1 protein was up-regulated in SK-MEL-2 cells transfected with Beclin1 plasmid;The invasion and migration ability of SK-MEL-2 cells overexpressed by Beclin1 decreased.