title: Methylation-associated silencing of miR-495 inhibit the migration and invasion of human gastric cancer cells by directly targeting PRL-3.
abstract: Phosphatase of regenerating liver-3 (PRL-3) is believed to be associated with cell motility, invasion, and metastasis. Our previous work found that PRL-3 is highly overexpressed in gastric cancer (GC) tissue with peritoneal metastasis and directly involved in the pathogenesis of GC peritoneal metastasis. Moreover, we further found that the down-regulation of endogenous miR-495 expression plays a causative role in over expression of PRL-3 in GC peritoneal metastasis. However, the molecular regulation mechanisms by which endogenous miR-495 expression is down-regulated and PRL-3 promotes GC peritoneal metastasis remain to be clearly elucidated. Some studies have shown that the promoter methylation is closely related to the miRNA gene expression. Therefore, in present study, based on our previous findings, we will analysis whether DNA methylation is a major cause of the down-expression of endogenous miR-495, which results in PRL-3 overexpression in GC peritoneal metastasis. Methylation specific PCR (MSP) and sodium bisulfite sequencing method (BSP) detected miR-495 gene promoter methylation status. We treated GC cell lines with 5-Aza-2'-deoxycytidine (5-Aza-dC) to make the gene promoter methylation inactivation. By treating with 5-Aza-dC the migration and invasion of GC cells were significantly inhibited. And the miR-495 was overexpressing, corresponds to the mRNA and protein levels of PRL-3 were reduced, the ability of invasion and metastasis was inhibited. This study suggest that miR-495 have tumor suppressor properties and are partially silenced by DNA hypermethylation in GC, will provide new strategies for prevention and treatment of GC peritoneal metastasis.
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China. Electronic address: lzr13@foxmail.com.
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Guoyang
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Daojiang
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China. Electronic address: jiezg123@126.com.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Jie
authors.affiliation: Key Laboratory of Clinical Pharmacology, Nanchang University, Nanchang, Jiangxi Province 330000, China. Electronic address: 13065192685@126.com.
authors.forename: Heping
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Jianbo
authors.initials: J
authors.lastname: Xiong
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Zhibiao
authors.initials: Z
authors.lastname: Le
authors.affiliation: Department of Gastrointestinal Surgery, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province 330000, China.
authors.forename: Shengxing
authors.initials: S
authors.lastname: Tan
mesh_terms: D001374:Azacitidine / Q000031:analogs & derivatives / Q000737:chemistry; D045744:Cell Line, Tumor; D002465:Cell Movement; D019175:DNA Methylation; D000077209:Decitabine; D020868:Gene Silencing; D016147:Genes, Tumor Suppressor; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013274:Stomach Neoplasms / Q000378:metabolism*
chemical_list: C577768:MIRN495 microRNA, human; D035683:MicroRNAs; D009363:Neoplasm Proteins; D000077209:Decitabine; C484730:PTP4A3 protein, human; D017027:Protein Tyrosine Phosphatases; D001374:Azacitidine
keywords: DNA methylation; Gastric carcinoma; PRL-3; miR-495