MicroRNA-200c-3p inhibits proliferation of nephroblastoma cells by targeting CCNE2.
10.12122/j.issn.1673-4254.2020.09.04
- Author:
Juan CAO
1
;
Liping SUN
2
;
Jianhong AN
2
;
Huan ZHANG
1
;
Xiaoxiao HE
1
;
Hong SHEN
2
Author Information
1. Department of Pathology, Shenzhen Children's Hospital, Shenzhen 518038, China.
2. Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515 China.
- Publication Type:Journal Article
- Keywords:
CCNE2;
microRNA-200c-3p;
nephroblastoma
- From:
Journal of Southern Medical University
2020;40(9):1246-1252
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To predict and verify the target gene of miR-200c-3p and evaluate the inhibitory effect of miR-200c-3p on the proliferation of nephroblastoma cells.
METHODS:The putative target genes of miR-200c-3p were predicted by bioinformatics approach. Nephroblastoma cell models with miR-200c-3p overexpression or knockdown were established in SK-NEP-1 and G401 cells with corresponding control groups. The expressions of CCNE2 in SK-NEP-1 and G401 cells in different groups were detected by RT-PCR and Western blotting. A luciferase reporter assay was used to determine the targeting relationship between miR-200c-3p and CCNE2. The effects of miR-200c-3p overexpression or knockdown on cell proliferation was detected by cell counting kit-8 (CCK-8) assay and soft agarose assay.
RESULTS:CCNE2 was one of the target genes of miR-200c-3p as predicted by bioinformatics methods. Transfection of the two nephroblastoma cell lines with miR-200c-3p mimic resulted in significantly lowered CCNE2 mRNA and protein expressions ( < 0.05). The results of dual-luciferase assay confirmed that miR-200c-3p bound to the 3'UTR of CCNE2. CCK-8 assay and soft agarose assay demonstrated that overexpression of miR-200c-3p significantly inhibited the proliferation of the nephroblastoma cells ( < 0.01), and knocking down miR-200c-3p in the cells produced the opposite effects.
CONCLUSIONS:miR-200c-3p overexpression inhibits the proliferation of nephroblastoma cells by down-regulating its target gene CCNE2.
