title: microRNA-199a-3p, DNMT3A, and aberrant DNA methylation in testicular cancer.
abstract: It was previously demonstrated that miR-199a was downregulated in testicular germ cell tumor (TGCT), probably due to hypermethylation of its promoter. Further study found that re-expression of miR-199a in testicular cancer cells (NT2) led to suppression of cell growth, cancer migration, invasion and metastasis. More detailed analyses showed that these properties of miR-199a could be assigned to miR-199a-5p, one of its two derivatives. The biological role of the other derivative, miR-199a-3p in TGCT, remains largely uncharacterized. In this report, we identified DNA (cytosine-5)-methyltransferase 3A (DNMT3A), the de novo methyltransferase, as a direct target of miR-199a-3p using a 3'-UTR reporter assay. Transient expression of miR-199a-3p in NT2 cells led to decrease, while knocking down of miR-199a-3p in a normal human testicular cell line (HT) led to elevation, of DNMT3A2 (DNMT3A gene isoform 2) mRNA and protein levels. In clinical samples, DNMT3A2 was significantly overexpressed in malignant testicular tumor, and the expression of DNMT3A2 was inversely correlated with the expression of miR-199a-3p. However, DNMT3A did not affect miR-199a expression in NT2 cells. Further characterization of miR-199a-3p revealed that it negatively regulated DNA methylation, partly through targeting DNMT3A. Overexpression of miR-199a-3p restored the expression of APC and MGMT tumor-suppressor genes in NT2 cells by affecting DNA methylation of their promoter regions. Our studies demonstrated the deregulation of miR-199a-3p expression in TGCT may provide novel mechanistic insights into TGCT carcinogenesis and suggested a potentially therapeutic use of synthetic miR-199a-3p oligonucleotides as effective hypomethylating compounds in the treatment of TGCT.
authors: Key Laboratory for Regenerative Medicine; Ministry of Education; School of Biomedical Sciences; Faculty of Medicine; The Chinese University of Hong Kong; Hong Kong, P.R. China.
authors: Bi-Feng
authors: BF
authors: Chen
authors: Key Laboratory for Regenerative Medicine; Ministry of Education; School of Biomedical Sciences; Faculty of Medicine; The Chinese University of Hong Kong; Hong Kong, P.R. China.
authors: Shen
authors: S
authors: Gu
authors: Key Laboratory for Regenerative Medicine; Ministry of Education; School of Biomedical Sciences; Faculty of Medicine; The Chinese University of Hong Kong; Hong Kong, P.R. China.
authors: Yick-Keung
authors: YK
authors: Suen
authors: Key Laboratory for Regenerative Medicine; Ministry of Education; School of Biomedical Sciences; Faculty of Medicine; The Chinese University of Hong Kong; Hong Kong, P.R. China.
authors: Lu
authors: L
authors: Li
authors: Key Laboratory for Regenerative Medicine; Ministry of Education; School of Biomedical Sciences; Faculty of Medicine; The Chinese University of Hong Kong; Hong Kong, P.R. China.
authors: Wai-Yee
authors: WY
authors: Chan
mesh_terms: D018236:Carcinoma, Embryonal / Q000378:metabolism*; D045744:Cell Line, Tumor; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D018239:Seminoma / Q000378:metabolism*; D013736:Testicular Neoplasms / Q000378:metabolism*
chemical_list: C000717967:DNMT3A protein, human; D035683:MicroRNAs; C531408:mirn199 microRNA, human; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A
keywords: DNA methylation; DNMT3A; miR-199a-3p; testicular cancer
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