Effect On MMPs Expression Of Prostate Cancer PC-3 Cell Lines By ～(60)Coγ-ray External Irradiation

Prostate cancer is the second leading cause of cancer deaths of men in the United States,and the vast majority of Pca deaths are related to metastatic disease. The incidence and mortality rates of the prostate cancer is increasing in our country with the life-span prolonging in recent years. The progress of prostate cancer is not only depend on the cell proliferation, but also closely related with the cellular biological behavior such as the migration and invasiveness.Unfortunately,there are few treatment options for patients with advanced Pca.Despite advances in androgen ablation therapies,the cancer will often progress to an androgen independent metastatic stage. Radiation therapy is the definitive treatment options for advanced Pca. OBJECTIVE(1) To study the relationship between the expression of MMP-2, MMP-9, EPO-R, CD31 of prostate cancer specimen and tumor grade,stage by immunohistochemical method.(2) To study the expression of MMP-2, MMP-9, EPO-R of the PC-3 cell lines (after irradiation with different dose of the γ-rays) by immunohistochemical method.(3) To study the expression of MMP-2, MMP-9, EPO-R of the PC-3 cell lines( after irradiation with different dose of the γ-rays ) by RT-PCR,FCM and Westernblottingjto investigate the capability of the migration, invasiveness of PC-3 cell befor and after irradiation. This study was aimed at investigating the role of MMP-2, MMP-9,EP0-R in prostate cancer,which expect to help the radiotherapy for the prostate cancer.METHODS(1)IHC was performed using the two-way method to study MMP-2, MMP-9,EPO-R,and CD31 expression of the prostate cancer surgical specimen.(2)IHC was performed using the two-way method to study MMP-2, MMP-9, and EPO-R expression of the cultured PC-3 cell lines( befor and after irradiation with different dose of the y-rays).(3)Cell proliferation was performed using MTT and clony formation; Assay of MMP-2 and MMP-9 proteinase were performed using FCM and Westernblotting; The gene expression of MMP-2 and MMP-9 was analyzed by RT-PCR; at the same time, Invasion of PC-3 cells in vitro was measured by the invasion of cells through Matrigel-coated transwell inserts(the cultured PC-3 cell lines,befor and after irradiation with 1 Gy,3 Gy,5 Gy dose γ-rays). RESULTS· IHC shown significant differences of MMP-2, MMP-9, CD31 and EPO-R expression bewteen Pea and BPH, and closely related with the prostate cancer pathologic grade, state and Gleason score,but showed no significant association between the amount of these factors labeling and the serum prostate particularity antigen( PSA).· PC-3 cell proliferation were significantly inhibited by the γ-rays(1Gy, 3Gy, 5Gy), the cell colony rate decreased following the increasing dose of ⁶⁰Co γ-rays.Matrigel Invasion Assay in vitro shown that sublethal irradiation promotes migration and invasion of PC-3 cell lines,· RT-PCR shown the gene expression of MMPs mRNA increased following the increasing dose of ⁶⁰Co y-rays, FCM and Western Blottings shown MMP-9 overexpression after irradiation with the γ- rays ( p<0.05),EPO- R overexpression in the 1 Gy and the 3 Gy groups.CONCLUSION· Our study suggests MMPs and EPO-R overexpression in the malignant prostate glands, MMPs may be a predictor of prostate cancer independent of disease stage, prostate-specific antigen(PSA), and Gleason score(GS).· Our study suggests that PC-3 cell migration and invasiveness enhanced by...