MAP9 Exhibits Protumor Activities and Immune Escape toward Bladder Cancer by Mediating TGF-β1 Pathway

Abstract

To investigate more potential targets for the treatment of human bladder cancer, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and high-content screening (HCS) analysis were performed, and microtubule-associated protein 9 (MAP9), which had the strongest proliferation inhibition from 809 downregulated genes, has been selected. MAP9 is responsible for bipolar spindle assembly and is involved in the progression of many types of tumors; however, its role in bladder cancer (BC) remains unknown. Expressive levels of MAP9 in BC tissues were determined through immunohistochemistry, and the clinical significance of MAP9 in BC was analyzed. Short hairpin ribonucleic acid- (ShRNA-) MAP9 was used to construct stable MAP9 knockdown BC cell lines. The proliferative abilities of MAP9 were measured through assays in vivo and in vitro, and the migrated and invasive abilities of MAP9 were analyzed via in vitro experiments. Quantitative reverse transcription PCR, western blotting, coimmunoprecipitation (Co-IP), and rescue assays were used to identify downstream targets of MAP9. MAP9 expression increased in the tumor tissues, and its increased level was negatively correlated with prognosis. Further, the loss of MAP9 caused decreased BC cell proliferation via inducing the growth 1/synthesis (G1/S) cell cycle arrest in vitro and slowed tumor growth in vivo. In addition, MAP9 silencing attenuated BC cell migration and invasion. Moreover, we found that the growth 1/synthesis (G1/S) cell cycle-related genes and the epithelial mesenchymal transition (EMT) marker levels decreased after silencing MAP9. Finally, we found that the transforming growth factor beta 1 (TGF-β1) pathway is activated as a mediator for MAP9 to regulate genes related to the G1/S cell cycle and EMT. MAP9 promotes BC progression and immune escape activity through the TGF-β1 pathway and is a potential novel target for therapies of BC.