signature=6f60ab606850e3d10348de6feacdc25c,MP36-08 MICRORNA-144-5P FUNCTIONS AS TUMOUR SUPPRESSOR TH...

摘要：

Introduction and Objective: Our recent study of microRNA (miRNA) expression signature in bladder cancer (BC) revealed that miR-144-5p expression was significantly reduced in BC tissues, suggesting miR-144-5p function as a tumour suppressive miRNA in BC cells. The aim of this study was to investigate the functional significance of miR-144-5p and to identify its regulated oncogenic genes in BC. Methods: Expression levels of miR-144-5p and its candidate target genes were evaluated in BC cell lines (T24, BOY, J82, UMUC, and KK47) and BC clinical specimens (60 BCs and 22 normal bladder epitheliums: NBEs) by qRT-PCR methods. Gain-of-function studies (cell proliferation, migration, invasion, apoptosis, and cell cycle assays) were performed using transfection of mature miR-144-5p into cancer cells. Genome-wide gene expression analysis and in silico analysis were applied to investigate molecular targets regulated by miR-144-5p in BC cells. A luciferase reporter assay was carried out to determine whether 3' UTR of target genes have actual biding sites for miR-144-5p. Overall survival (OS) of the BC patients was evaluated using the Kaplan-Meier method. Results: The expression levels of miR-144-5p was significantly reduced in BCs and BC cell lines compared to NBEs (P<0.0001). Restoration of miR-144-5p significantly inhibits cancer cell proliferation, migration, and invasion. Flow cytometry analysis revealed that G1 arrest was observed in miR-144-5p transfectants. Four cell cycle-related genes (CCNE1, CCNE2, CDC25A, and PKMYT1) were identified for direct target genes of miR-144-5p by microarray and in silico studies, and luciferase reporter assays. Kaplan-Meier analysis showed that the patients with high CCNE1 and CCNE2 expressions had significantly shorter OS probability than those with low expressions of the genes (P=0.025 and 0.032). Conclusions: Downregulation of miR-144-5p was frequent event of BC cells and its function as a tumour suppressor via targeting cell cycle-related genes such as CCNE1 and CCNE2. Overexpression of these genes might be contributed to progression of BC oncogenesis. Elucidation of tumour-suppressive miR-144-5p regulated molecular pathways and targets in BC cells could provide new information on potential diagnostic markers and therapeutic targets in BC.

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