Effect of lncRNA CDKN2B-AS1 on malignant biological behaviors of melanoma B16-F10 cells
10.3872/j.issn.1007-385x.2020.01.004
- VernacularTitle:lncRNACDKN2B-AS1对黑色素瘤B16-F10细胞恶性生物学行为的影响
- Author:
DENG Lili
1
;
SUN Sujiao
2
;
LIU Yan
1
;
CHEN Yan
3
;
CHEN Xing
3
Author Information
1. Department of Dermatology, Chengdu Second People's Hospital
2. Department of Aesthetics, the First Affiliated Hospital of Dali University,
3. Department of Dermatology,Affiliated Hospital of North Sichuan Medical College,
- Publication Type:Journal Article
- Keywords:
lncRNACDKN2B-AS1;
miR-7-5p;
melanoma;
B16-F10 cell;
proliferation;
migration;
invasion
- From:
Chinese Journal of Cancer Biotherapy
2020;27(1):25-30
- CountryChina
- Language:Chinese
-
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 mRNAexpression of miR-7-5p, and protein expressions of cleaved caspase-3 and E-cadherininsh-CDKN2B-AS1groupandmiR-7-5pmimic group were significantly up-regulated (all P<0.05), whiletheproteinexpressionsofKi67,N-cadherin,andTwist1weresignificantlydown-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 malignant biological behaviors of melanoma B16-F10 cells.
- Full text:20200104.pdf
