| Objective The study was designed to evaluate the effect of the selective cyclooxygenase 2 inhibitor Celecoxib and the endothelin A receptor antagonist BQ123, alone and incombination, on the growth and cell apoptosis of the human prostate cancer cell line PC3 and PC3M, and to investigate the possible mechanisms ivolved in these effects.Methods In the first part of this study, the viability of PC3 and PC3M cells with Celecoxib and/or BQ123 were measured by MTT assay. The migration distances were measured by wound healing assay. The effects on the migration and invasion ability of these two cell lines with Celecoxib and/or BQ123 were determined by the cell migration assay in Transwell model. In the second part of this study the Annexin V-FITC/PI assay was used to investigate the apoptosis of the cells. The relationship between Celecoxib and/or BQ123 on PC3 and PC3M and the cell cycle was observed by cell cycle assay with flow cytometry. All the information was analyzed by SPSS vl3.0 software. P value of <0.05 was considered statistically significant.Results The MTT assay showed that accompanied with the increasing concentration of Celecoxib and its affected time it, the inhibitory ratio increased in a dose-dependant and time-dependant fashion (p<0.05). The cells were all dead in 400μM which the inhibitory ratio was 100%. The IC50 in 24 hour which the Celecoxib inhibited PC3 and PC3M cells growth was 50μM. Accompanied with the increasing concentration and affected time of BQ123, the inhibitory ratio also increased in a dose-dependant and time-dependant fashion (p<0.05). Used the concentration less than IC50 of Celecoxib in combination with BQ123 on the cancer cells, the inhibition was correspond to the Celecoxib alone on the cells in the concentration of 100μM and 200μM. It indicated that the inhibition effect which exerted Celecoxib combined with BQ123 on the cells would be more powerful than exerted them respectively.The wound healing assay showed that the migration distances of the 100μM Celecoxib group were all less than the control group accompanied with the increasing concentration and the affected time(p<0.05). The migration distances of the group Celecoxib in 100μM combined with BQ123 in 1μM were less than which of the group BQ123 in the same concentration, but not significantly decreased compared with the group which Celecoxib in the same concentration(p>0.05). It indicated that the combination and the Celecoxib in 100μM could inhibit the cell migration ability of the cancer cells(p<0.05), and the inhibition effect which exerted Celecoxib combined with BQ123 on the cells would be more powerful than exerted BQ123 in the same concentration respectively(p<0.05). There was statistical significance in the migration distances in 72 hours when combination compared with Celecoxib in the same concentration respectively(p<0.05).The cell migration assay showed that the numbers PC3 and PC3M cells invaded to the inferior chamber which the 100μM Celecoxib group, 1μM BQ123 group and 100μM Celecoxib in combination with 1μM BQ123 group compared to the control group were significantly decreased. The numbers of the combination group were decreased singnificantly compared to the Celecoxib or BQ123 alone group. It indicated that Celecoxib in 100μM and 1μM BQ123, alone or in combination, could inhibit the cell migration and invasion ability of the cancer cells(p<0.05), and the inhibition effect which exerted Celecoxib combined with BQ123 on the cells would be more powerful than exerted BQ123 in the same concentration respectively(p<0.05).The Annexin V-FITC/PI staining assay showed that Celecoxib and BQ123 all could induce the apoptosis of PCa cells. The induction ability of BQ123 in 1μM was weak. The induction effect of Celecoxib in 100μM was significantly obvious. The induction effect of the combination group was better than exerting them in the same concentration respectively.The cell cycle assay showed that the apoptosis ratio of PC3 and PC3M cells, which exerted by Celecoxib in 100μM and 100μM Celecoxb in combination with 1μM BQ123, in G0/G1 phase increased significantly compared to the control group. The ratio in S phase decreased significantly compared to the control group(p<0.05). It indicated that the cell cycle was blocked by the Celecoxib and/or Celecoxib combined with BQ123 in G0/G1 phase. The apoptosis ratio in every phase of the BQ123 group compared to the control group was not changed significantly(p>0.05). The apoptosis ratio of combination group compared to the 100μM Celecoxib group was not changed significantly(p>0.05). So BQ123 was not significantly affect to the cell cycle of the PCa cells.
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