RESUMEN
PURPOSE: GRHL2 played important roles in different cancers. In this study, we aimed to investigate the roles of GRHL2 in bladder cancer. METHODS: The immunohistochemistry assay was performed to detect the expression of GRHL2 in bladder cancer tissues and adjacent noncancerous tissues and the expression levels of GRHL2 and zinc finger E-box binding homeobox (ZEB1) mRNA in tissues were determined by qRT-PCR. In addition, qRT-PCR and Western blotting were applied to detect the expression levels of GRHL2 and ZEB1 in bladder cancer cell lines (RT4, BIU-87, 5637, T24) and immortalized human bladder epithelial cell line (SV-HUC-1). The cell models with up-regulated and down-regulated expression of GRHL2 were constructed using bladder cancer cell lines T24 and 5637 to investigate the underlying roles of GRHL2 on the proliferation, migration, invasion and EMT process of bladder cancer cells. After that, cell proliferation was evaluated by CCK8 assay, cell cycle assay and colony formation assay. Transwell assay and wound healing assay were performed to determine the invasion and migration ability of the bladder cancer cells. The expressions of epithelial-mesenchymal transition (EMT) related proteins (E-cadherin, Vimentin, Slug and Snail) were assessed by Western blot analysis. Moreover, ZEB1 and GRHL2 were co-transfected into T24 and 5637 cells and their effects on EMT process and invasive capacity of cells were examined. RESULTS: The expression of GRHL2 was down-regulated in bladder cancer tissues and human bladder cancer cell lines compared with the normal bladder tissues and immortalized human bladder epithelial cell line. Besides, down-regulation of GRHL2 improved the proliferation ability of bladder cancer cells and promoted the EMT process through up-regulation of ZEB1. The overexpression of ZEB1 partially reversed the inhibitory effect of GRHL2 on EMT. CONCLUSION: GRHL2 acts as an anti-oncogene to regulate bladder cancer cell proliferation and inhibit EMT by targeting ZEB1. This study may provide a theoretical basis for further research.
RESUMEN
The function of miR-186 in the progression of renal cell carcinoma (RCC) remains poorly investigated. Our study aims to identify the molecular mechanism underlying miR-186-regulated proliferation, migration and invasion of RCC. Firstly, our data confirmed that miR-186 was significantly reduced and CDK6 was obviously increased in RCC tissues and cells. MiR-186 or CDK6 was associated with advanced TNM stage, lymph node metastasis and poor prognosis. MiR-186 significantly inhibited cell proliferation, migration, invasion and in vivo tumor growth, induced apoptosis, and blocked cell cycle progression in G0/G1 phase. MiR-186 also induced Bax expression and inhibited the expressions of Bcl-2, cyclin D1 and epithelial-mesenchymal transition (EMT)-related genes. Additionally, CDK6 expression was downregulated by miR-186 via binding to its 3'-untranslated region (3'-UTR). Moreover, ectopic expression of CDK6 could partially abrogate the inhibitory effect of miR-186. In conclusion, miR-186 suppresses proliferation, migration and invasion of RCC by inhibiting CDK6 expression.
Asunto(s)
Carcinoma de Células Renales/genética , Quinasa 6 Dependiente de la Ciclina/genética , Quinasa 6 Dependiente de la Ciclina/metabolismo , Neoplasias Renales/genética , Neoplasias Renales/patología , MicroARNs/fisiología , Regiones no Traducidas 3'/genética , Apoptosis/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Progresión de la Enfermedad , Regulación hacia Abajo/genética , Transición Epitelial-Mesenquimal/genética , Expresión Génica/genética , Humanos , Metástasis Linfática/genética , Metástasis Linfática/patología , Invasividad Neoplásica/genética , Fase de Descanso del Ciclo Celular/genéticaRESUMEN
PURPOSE: MicroRNAs have recently reported playing a vital role in the development of cancers. However, the role of miR-592 in renal cell carcinoma (RCC) has not been explored. In this study, the potential role of miR-592 was investigated in RCC. PATIENTS AND METHODS: The expression of miR-592 was evaluated in RCC tissues and cell lines using qRT-PCR assays. The Kaplan-Meier analysis and Cox proportional hazards model analysis was used to analyze the prognostic value of miR-592 in RCC. The effects of miR-592 on cell proliferation, migration, and invasion were determined by cell counting kit-8 (CCK-8) and Transwell assays in vitro. RESULTS: The results showed that miR-592 was significantly increased both in RCC tissues and cell lines. Overexpression of miR-592 was significantly associated with lymph node metastasis, TNM stage, and poor overall survival. And functional studies in two RCC cell lines (786-O and Caki-1) have shown that overexpression of miR-592 promoted cell proliferation, migration, and invasion, while silence of miR-592 inhibited cell proliferation, migration, and invasion. SPRY2 was a direct target of miR-592. CONCLUSION: Overall, overexpression of miR-592 may be a prognostic biomarker and therapeutic strategy for patients with RCC, which is correlated with the progression of RCC.