YAP1 Promotes EMT Phenotype Of PCa And Its Mechanism Research

Objective:We focus on the effect of YAP1 with prostate cancer EMT phenotype and do some preliminary research on its mechanism through detecting YAP1 and EMT markers of prostate cancer patients with various disease stages and doing related cell and mice experiments. Methods:1, From BPH to CRPC, measuring and comparing the protein level of YAP1, E-Cadherin and N-Cadherin.2, For cell culturing part, firstly, after silencing YAP1 gene, conduct wound healing assay and observe cell migration.secondly, comparing the differences of YAP1, E-Cadherin and N-Cadherin in PCa cells after silencing or overexpressing YAP1.thirdly, comparing the differences of AR, YAP1 and P-Smads between cytoplasm and nucleus after the treatment of DHT or Enzalutamide.finally, observing the change of YAP1 expression and cell phenotype after TGF-? treatment.3, For mice experiments part, we verify the above trials in mice and at the same time study the function of Verteporfin in inhibiting xenograft prostate tumors growth, YAP1 expression and tumor EMT phenotype. Results:1, IHC results show that expression level of YAP1 and N-Cadherin increase gradually from BPH, low Gleason score ADPC, high Gleason score ADPC, ADT clinical stage to CRPC.However, E-Cadherin presents a reducing trend in the five disease stages.2, Wound healing assay indicates that knockdown of YAP1 in C4-2 shortens the migration distance significantly.Likewise,E-Cadherin increases and Vimentin decreases distinctly after silencing YAP1 in C4-2 cell.On the contrary, Vimentin increases and E-Cadherin decreases distinctly after YAP1 overexpression in C4-2 cellline.There is no observably difference of AR, YAP1 and P-Smads in cytoplasmic protein after DHT or enzalutamide treatment in LNCaP.Surprisingly, we find that AR decreases variously, at the same time, YAP1 and P-Smads increase greatly in nucleus after the above administration. Besides, we find that TGF-? can promote the transcription of YAP1 gene with statistical significance.3, we find that C4-2 xenografts with YAP1 knockdown do not decrease without obvious metastasis in BALB/c mice. Verteporfin can shrink C4-2 xenograft tumor sizes than DMSO and IHC datas show that expression level of YAP1 and N-Cadherin decrease remarkably, otherwise, E-Cadherin content increases.Brdu assay shows that tumor proliferates more fiercely in VP drug group than control group with statistical siginificance. Along with castration surgery of BALB/c mice with LNCaP xenograft tumors, YAP1 and N-Cadherin proteins express progressively but the level of E-Cadherin is reducing gradually. Conclusions:Our study preliminarily confirms that YAP1 protein can promote PCa EMT phenotype change and enhance cancer cell migration capacity. Its mechanism may be that in the wake of dysfunction of AR or decrease of AR in nucleus, YAP1 activates gradually and promotes Smads phosphorylation therefore they translocate into nucleus collaboratively. Besides, classic TGF-? can also reinforce the above function of YAP1. Furthermore, verteporfin can inhibit the function of YAP1 in promoting PCa EMT change and has a relatively safe quality. In brief, through initial research and analysis, we find a probable molecular system, however, we have not made out a clear and complete signaling pathway or detailed EMT development mechanism of PCa. But anyway we still insist that YAP1 maybe have a fine development trend in the aspects of both prevention and treatment of CRPC or even mCRPC.