https://orcid.org/0000-0001-5241-9506
Figures & data
Figure 1. LncRNA SNHG8 level is enhanced in melanoma tissues and cells
(A) RT-qPCR was used to measure the expression level of lncRNA SNHG8 in melanoma tissues.(B) RT-qPCR showed the level of lncRNA SNHG8 in melanoma cells (A375, A875, M14, and SK-MEL-5) and normal epidermal melanocytes (HEMa-LP).(C) Kaplan–Meier assays were carried out to evaluate the association between lncRNA SNHG8 levels and overall survivals.*P < 0.05; **P < 0.01
Figure 2. LncRNA SNHG8 is activated by YY1
(A) The predicted sites of YY1-binding in lncRNA SNHG8 promoter were obtained from JASPAR.(B and C) RT-qPCR was used to measure YY1 expression in melanoma tissues and cells.(D) ChIP assay confirmed the enrichment of lncRNA SNHG8 promoter in the beads conjugated with anti-YY1 or anti-IgG.(E) Luciferase reporter assay revealed the interaction of YY1 with lncRNA SNHG8 promoter.(E) expression of YY1 was confirmed by RT-qPCR in A375 and A875 cells transfected with shYY1, shNC, pcDNA3.1 or pcDNA3.1/YY1.(G) RT-qPCR showed the level of lncRNA SNHG8 after the overexpression or knockdown of YY1.*P < 0.05; **P < 0.01;***P < 0.001
Figure 3. LncRNA SNHG8 knockdown suppresses melanoma cell viability and metastasis
(A) RT-qPCR was performed to confirm the transfection efficiency of sh-SNHG8 in the A375 and A875 cells.(B) Cell viability (C) proliferation, (D) cell invasion, and (E) cell migration was investigated in the A375 and A875 cells following transfection with sh-NC or sh-SNHG8 using CCK-8, colony formation, Matrigel, and wound healing assays, respectively.**P < 0.01
Figure 4. LncRNA SNHG8 binds to miR-656-3p
(A) The binding sequences of lncRNA SNHG8 and miR-656-3p, as predicted by the bioinformatic databases.(B) RT-qPCR was performed to confirm the transfection efficiency of miR-656-3p mimics in A375 and A875 cells.(C) Luciferase reporter assay verified the binding of miR-656-3p to SERBP1 in A375 and A875 cells.(D and E) RT-qPCR was used to measure miR-656-3p expression in melanoma tissues and cells.(F) Pearson’s correlation analysis analyzed the negative relation between miR-656-3p and lncRNA SNHG8 in our cohort.(G) RT-qPCR showed miR-656-3p expression in melanoma cells transfected with sh-NC or sh-SNHG8.*P < 0.05; **P < 0.01
Figure 5. miR-656-3p directly targeted SERBP1
(A) The binding sequences of miR-656-3p and SERBP1 were predicted by the bioinformatic databases.(B) The biding site of SERBP1 to miR-656-3p was validated using a luciferase reporter assay in A375 and A875 cells.(C and D) RT-qPCR was used to measure SERBP1 expression in melanoma tissues and cells.(E) Pearson’s correlation analysis analyzed the negative relation between SERBP1 and miR-656-3p or lncRNA SNHG8.(F) RT-qPCR was used to analyze the expression level of SERBP1 in the melanoma cells following transfection with sh-NC, sh-SNHG8, sh-SNHG8+ miR-656-3p inhibitor.*P < 0.05; **P < 0.01
Figure 6. miR-656-3p impedes melanoma cell viability, migration, and invasion via SERBP1
(A and B) RT-qPCR and Western blotting showed SERBP1 level in A375 cells transfected with NC mimics, miR-656-3p mimics, pcDNA3.1, SERBP1, and miR-656-3p mimics + SERBP1.(C and D) CCK-8 and colony formation experiments showed cell viability in A375 cells transfected with NC mimics, miR-656-3p mimics, pcDNA3.1, SERBP1, and miR-656-3p mimics + SERBP1.(E) Wound healing assay showed cell migration in A375 cells transfected with NC mimics, miR-656-3p mimics, pcDNA3.1, SERBP1, and miR-656-3p mimics + SERBP1.(F) Matrigel assay showed the invasion ability of A375 cells transfected with NC mimics, miR-656-3p mimics, pcDNA3.1, SERBP1, and miR-656-3p mimics + SERBP1.*P < 0.05; **P < 0.01.
