miR-145 inhibits invasion of bladder cancer cells by targeting PAK1

Bo Kou, Yang Gao, Chong Du, Qi Shi, Shan Xu, Chen-Qing Wang, Xinyang Wang, Dalin He, Peng Guo
Urologic Oncology 2014, 32 (6): 846-54

OBJECTIVES: MicroRNAs play important roles in cancer. In many cancers, miR-145 acts as a tumor suppressor, and it is down-regulated in bladder cancer. In the present study, we explored the modulation of oncogenic gene PAK1 by miR-145 in bladder cancer.

MATERIAL AND METHODS: Expression of miR-145 was detected in bladder cancer tissues and cell lines by quantitative real-time polymerase chain reaction. Through the bioinformatics approach, PAK1 has been predicted to be a direct target of miR-145 and was confirmed by the PAK1 messenger RNA 3'-untranslated region luciferase activity assay. To investigate whether miR-145 regulates PAK1 expression, it was overexpressed in J82 and T24 bladder cancer cells. In 10 paired bladder normal and tumor tissues, we determined the relationship between miR-145 and PAK1 through quantitative real-time polymerase chain reaction and western blot. By using transwell invasion assay and western blotting analysis, we investigated the effects of miR-145 and PAK1 on bladder cancer cell invasion and expression of invasion marker genes.

RESULTS: The level of miR-145 decreases and PAK1 protein expression up-regulates in bladder cancer tissue, as compared with the paired normal bladder tissue. Moreover, miR-145 directly targets PAK1 in bladder cancer cells. The level of miR-145 negatively correlates with PAK1 protein expression in bladder cancer. In addition, PAK1 promotes invasion and enhances the expression and activity of MMP-9, whereas miR-145 inhibits bladder cancer cell invasion and expressions of PAK1 and MMP-9.

CONCLUSIONS: Our results indicate that miR-145 inhibits bladder cancer cell invasion, at least partly through targeting PAK1. Restoration or replacement of miR-145 could be an efficient approach to inhibit PAK1 and bladder cancer development in the tumor therapy.

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