MicroRNA-30a-5p Suppresses Malignant Progression and is Associated with Clinical Diagnosis and Prognosis of Urothelial Carcinoma of the Bladder by Modulating Proliferating Cell Nuclear Antigen Clamp Associated Factor

Abstract

Background and Objectives: The diagnostic and therapeutic approaches to urothelial carcinoma of the bladder (UCB) have their limitations. The aim of this study was to explore whether microRNA-30a-5p (miR-30a-5p)/proliferating cell nuclear antigen clamp associated factor (PCLAF) can serve as biomarkers for diagnosis, predicting patient prognosis, and UCB treatment. Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine miR-30a-5p and PCLAF expressions in clinical UCB tissues and human UCB cell lines. Western blotting (WB) and immunohistochemistry (IHC) were used to quantify the levels of PCLAF protein in human UCB tissues and xenograft tumor tissues, respectively. The link between miR-30a-5p and PCLAF mRNA/protein level in human UCB tissues was examined a Spearman correlation analysis. miR-30a-5p/PCLAF’s function on human UCB cell proliferation was examined, using the cell counting kit-8 (CCK-8)/5-bro’o-2’-dexoyuridine (BrdU)/3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazol-ium bromide (MTT)/colony formation experiments, on migration with Transwell/Wound healing assays, and on invasion with Transwell assay. Tumor growth and cell metastasis models were established to evaluate the roles of miR-30a-5p/PCLAF on human UCB tumor development and metastasis, respectively. Results: MiR-30a-5p and PCLAF were significantly underexpressed and upregulated, respectively, in human UCB tissues and cells, and their expression patterns significantly and negatively correlated in human UCB tissues. MiR-30a-5p upregulation significantly suppressed while miR-30a-5p knockdown significntly stimulated cell proliferation, migration and invasion, tumor development and metastasis in human UCB, by regulating PCLAF expression. Moreover, downregulated miR-30a-5p or upregulated PCLAF expression were significantly associated with increased progression and poor prognosis of UCB sufferers. Conclusions: MiR-30a-5p significantly suppressed UCB malignant progression, by regulating PCLAF. MiR-30a-5p and PCLAF have a potential to be the possible biomarker for UCB’s diagnosis and prognosis.