The TGF-?/EGFR Signal Pathway Promoting The Process Of EMT In Prostate Cancer Cell

Objective1. To study the expression of TGF-? in the tissues of androgen dependent prostate cancer and CRPC patients, its serum values and analyze its clinical significance.2.To investigate the role of TGF-?/EGFR pathway in the EMT process of prostate cancer cells. MethodsWe used the immunohistochemical staining technique to detect the expression of TGF-? in different prostate tissues and Luminex Liquichip to detect the level of TGF-? in the serum of these two groups of patients and analyzed their relationship with clinicpathological factors.We then applied the Western blot and RT-qPCR technique to detect the protein expression and mRNA level of TGF-?, EGF, EGFR, E-cadherin and Vimentin in LNCaP, C42, DU145 and PC3 prostate cancer cells. We chose the LNCaP and C42 prostate cancer cells as the experimental objects, treating them with 50ng/ml TGF-?. After 24 h treatment, Western blot and RT-qPCR were applied to detect the mRNA quantity and protein level of E-cadherin and Vimentin, We also detected the expression of p-EGFR protein. Transwell was used to assess invasion of LNCaP and C42 cells treating with TGF-? and Wound Healing assay was used to assess migration of the cells.We summarized all the experimental data and used the SPSS 19.0 statistical software to analyze the data. Results1. The immunohistochemical staining result showed that the expression of TGF-? in CRPC tissues was higher than that in prostate cancer tissues(P<0.05) and the level of TGF-? in the serum of CRPC patients was also higher than that of prostate cancer patients(P<0.05). 2. Following the order of LNCaP, C42, DU145, PC3, the mRNA and protein level of TGF-? raised gradually, while the level of EGF decreased gradually. The mRNA and protein level level of E-cadherin is very high in LNCaP and C42 cell lines, while the mRNA level of Vimentin is very low in these two cell lines, we did not detect the expression of Vimentin protein in LNCaP and C42 cell lines. The mRNA and protein level of Vimentin is very high in DU145 and PC3 cell lines, while the mRNA level of E-cadherin is very low in these two cell lines, the protein level of E-cadherin is very low in DU145 cell lines and we did not detect the expression of E-cadherin protein in PC3 cell lines. 3. The results of Western bolt demonstrated that the protein level of E-cadherin decreased while the protein level of Vimentin increased significantly in the LNCaP and C42 cell lines treated with TGF-?.We also found that treating the cells with TGF-? can increase the migration and invasion abilities of LNCaP and C42 cell lines greatly, suggesting its role in the EMT. ConclusionOur findings suggested that the expression of TGF-? is correlated with the progression of CRPC development and TGF-?/EGFR pathway can result in EMT in LNCaP and C42 cell lines, suggesting that TGF-? may serve as a promising biomarker and therapeutic target for prostate cancer.