TSPYL2 RNA Hypomethylation Inhibits Prostate Cancer Cell Migration and Proliferation

Abstract

Background: Testis-specific protein Y-encoded-like 2 (TSPYL2) has been identified as a tumor suppressor on the X chromosome whose hypomethylation is regarded as a biomarker of uterine leiomyomas. Its methylation status in prostate cancer (PCa) remains unclear. The purpose of this study was to investigate the methylation status of TSPYL2 in PCa and its role in the migration and proliferation of PCa cells. Methods: Following the required transfection with short hairpin RNA-methyltransferase-like 3 (shRNA-METTL3) and TSPYL2 overexpression plasmid, both quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot assays were adopted to measure the expressions of METTL3 and TSPYL2. The content of N6-methyladenosine (m6A) was measured through RNA immunoprecipitation (RIP). Whether TSPYL2 RNA was methylated in PCa cells was validated by RIP-qPCR. The level of TSPYL2 RNA methylation was determined through Methylated (Me) RIP-qPCR. Scratch and colony formation assays were separately adopted to evaluate migration and proliferation, respectively. Results: TSPYL2 expression was down-regulated (p < 0.001) while METTL3 expression was up-regulated (p < 0.001) in PCa cells (LNCaP and PC3). The expression level of TSPYL2 was down-regulated in prostate adenocarcinoma (PRAD) tumor samples (p < 0.05). m6A content was high in LNCaP and PC3 cells (p < 0.01). TSPYL2 possessed METTL3-related methylation in PCa cells. METTL3 positively regulated TSPYL2 methylation in PCa cells. Both shRNA-METTL3 and TSPYL2 overexpressions decreased the migratory and proliferative abilities of LNCaP and PC3 cells (p < 0.001). Conclusions: TSPYL2-mediated RNA hypomethylation suppresses migration and proliferation of PCa cells.