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Effect of lncRNA CDKN2B-AS1 on malignant biological behaviors of melanoma B16-F10 cells
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Abstract:
Objective: To investigate the effect of long non-coding RNA CDKN2B antisense RNA 1 (CDKN2B-AS1) on malignant biological behaviors of melanoma B16-F10 cells by targeting miR-7-5p. Methods: Melanoma B16-F10 cells were chosen for this study.shRNA CDKN2B-AS1 vector was constructed and transfected into B16-F10 cells. The experimental cells were divided into control group, sh-CDKN2B-AS1 group, miR-7-5p mimic group and miR-7-5p inhibitor group. The expression level of CDKN2B-AS1 mRNA in the transfected B16-F10 cells was detected by RT-PCR; the number of clone formation and the proliferation ability of the cells were detected by Clone formation assay and MTT assay; and the migration and invasion ability of the cells were detected by Scratch-healing assay and Transwell assay. The targeting relationship between CDKN2B-AS1 and miR-7-5p was detected by Luciferase reporter gene assay. The mRNA expression of miR-7-5p and protein expressions of Ki67, cleaved caspase-3, E-cadherin, N-cadherin and Twist1 in B16-F10 cells after transfection with miR-7-5p mimics/inhibitor were detected by RT-PCR and Western blotting, respectively. Results:Compared with the control group, the expression level of CDKN2B-AS1 mRNA in B16-F10 cells of sh-CDKN2B-AS1 group was significantly decreased (P<0.01); the proliferation, migration and invasion ability of cells were significantly decreased (all P<0.01).Luciferase reporter gene assay showed that CDKN2B-AS1 directly targeted miR-7-5p. The mRNA expression of miR-7-5p, and protein expressions of cleaved caspase-3 and E-cadherin in sh-CDKN2B-AS1 group and miR-7-5p mimic group were significantly up-regulated (all P<0.05), while the protein expressions of Ki67, N-cadherin, and Twist1 were significantly down-regulated (all P<0.05). Conclusion:CDKN2B-AS1 targets miR-7-5p to promote the development of melanoma, and interfering with CDKN2B-AS1 can inhibit the malig‐nant biological behaviors of melanoma B16-F10 cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No.81560697)
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