miR-5100 mediates migration and invasion of melanomatous cells in vitro via targeting SPINK5

Abstract

The mechanism of miR-5100 and his target gene spink5 in the development of melanoma are rarely reported. Many data from targetscan and oncomine data base suggest that miR-5100 may regulate spink5 and mediate the biological behavior of melanoma. In this study, we used miR-5100's agomir and antagomir to interfere with the A375 cell line. Experimental data indicate that miR-5100 Agomir significantly reduces the expression of SPINK5, and miR-5100 Antagomir substantially increases the expression of SPINK5. Wound Healing assay showed that Compared with the control group, the healing distance of the SPINK5 inhibition group was shortened considerably, and the healing range of the SPINK5 overexpression group was substantially longer. Wound Healing assay showed that compared to the control group, migration of A375 cell line decreased during wound healing on high mRNA expression levels of SPINK5 and vice versa. Transwell assays further demonstrated that SPINK5 knockdown by Agomir significantly enhanced while SPINK5 overexpression markedly suppressed the invasion of A375 cells. Transwell assays also demonstrated that SPINK5 knockdown by Agomir significantly increased. In contrast, SPINK5 overexpression markedly suppressed the invasion of A375 cells. Western blot analysis showed that after overexpression of miR-5100, the expression of spink5 decreased, and the decrease of spink5 significantly promoted the expression of ERK1/2, p-ERK1/2, and p-AKT; the expression of spink5 increased after inhibiting the expression of miR-5100, and the expression of spink5 significantly inhibited ERK1/2, p-ERK1/2, p-AKT expression. Wound healing assay showed that compared to the control group, migration of A375 cell line decreased during wound healing on the MK-2206 group and SCH772984 group after using MK-2206, a p-Akt inhibitor, SCH772984, an ERK1/2 and p-ERK1/2 Inhibitor. Transwell assays suggest that compared with the control group, the invasive ability of groups MK-2206 and SCH772984 was significantly reduced. In conclusion, miR-5100 mediates migration and invasion of melanomatous cells in vitro via targeting SPINK5.