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06 December 2015 : Laboratory Research

Down-Regulation of miR-148a Promotes Metastasis by DNA Methylation and is Associated with Prognosis of Skin Cancer by Targeting TGIF2

Yanli TianABC, Wei WeiCDE, Li LiDE, Rongya YangEFG

DOI: 10.12659/MSM.894826

Med Sci Monit 2015; 21:3798-3805

Abstract

BACKGROUND: MicroRNAs (miRNA) dysregulation has been considered to be significantly related to the occurrence and development of cancers. Several studies had proved that DNA methylation is an important cause of the abnormal expression of miRNAs. The purpose of this study was to investigate the methylation status of miR-148a and its effects on the metastasis and prognosis of skin cancer, as well as the interaction with TGIF2 gene.

MATERIAL AND METHODS: According to the qRT-PCR analysis, the expression of miR-148a was down-regulated in tumor tissues compared with the adjacent tissues and healthy controls (P<0.05). In vitro cell metastasis assay revealed that miR-148a could inhibit cell metastasis and its down-regulation promoted metastasis. Luciferase reporter assay found that TGIF2 gene was a target gene and its expression was suppressed by miR-148a in skin cancer.

RESULTS: Methylation-specific PCR demonstrated that DNA methylation rate of miR-148a was higher in tumor tissues than in adjacent tissues and healthy tissues (P<0.05). miR-148a expression was proved to be epigenetically regulated after the demethylation of it by 5-aza-20-deoxycytidine treatment and qRT-PCR analysis. miR-148a methylation was significantly influenced by many clinicopathologic characteristics such as age (P=0.000), pathological differentiation (P=0.000), and lymph node metastasis (P=0.000). Besides, Kaplan-Meier analysis showed patients with miR-148a methylation lived shorter than those without that (P<0.001). Cox regression analysis manifested that miR-148a methylation (HR=0.053, 95CI%=0.005–0.548, P=0.014) could be serve as an independent prognostic marker for skin cancer.

CONCLUSIONS: Taken together, the expression of miR-148a was regulated by DNA methylation and targeted by TGIF2. Its methylation may be a potential prognostic indicator in skin cancer.

Keywords: Epigenesis, Genetic, Down-Regulation, DNA Methylation, Gene Knockdown Techniques, Homeodomain Proteins - genetics, MicroRNAs - genetics, Neoplasm Metastasis - genetics, Prognosis, RNA, Messenger - genetics, Repressor Proteins - genetics, Skin Neoplasms - pathology

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750