| The incidence of prostate cancer in China has increased year by year,and the proportion of patients with advanced has increased.There are more and more cases of hormone-resistant prostate cancer after endocrine therapy.At present,the common treatment for hormone-resistant prostate cancer is paclitaxel-based chemotherapy.However,many patients with hormone-resistant prostate cancer will progress with this treatment and die within 3 years,indicating hormone resistance.There is no good treatment for prostate cancer.Studies have shown that histone deacetylase inhibitors combined with chemotherapy or radiotherapy may become a way to treat tumors.The combination of paclitaxel and histone deacetylase inhibitors to treat hormone-resistant prostate cancer may also be an advantageous anti-tumor regimen.Therefore,it is meaningful to study the mechanism of histone deacetylase inhibitors against prostate cancer.In this paper,a three-part experiment was conducted to study the Akt/Fox03a signaling pathway of histone deacetylase inhibitor suberic anilide hydroxamic acid(SAHA)against hormone-resistant prostate cancer cells DU-145 and PC-3.Part I Effect of SAHA on proliferation,growth and apoptosis of prostate cancer DU-145 and PC-3METHODS:DU-145 and PC-3 cells were tested with different concentrations of SAHA.DU-145 was treated with 0,15 25 4,8,16,32 ?M,and PC-3 was treated with 0,0.25,0.5,1,5,10,15?M treatment,MTT method to measure the effect of SAHA on the viability of cells DU-145?PC-3.After PI staining and Annexin-V-FITC/PI staining,the effect of SAHA on prostate cancer cell cycle and apoptosis was detected by flow cytometry.Results:According to the IC50 value calculated by MTT,different concentrations were selected for subsequent experiments.The concentration of DU145 cells was 1,3,9?M,and the concentration of PC-3 cells was 0.5,2,8 ?M.The increase of SAHA concentration in the sub-GO/G1 phase tumor cells was statistically significant(P<0.05);the proportion of G2/M phase cells was increased and statistically significant(P<0.05).The percentage of apoptosis increased with the increase of SAHA concentration and was statistically significant(P<0.05).Part ? SAHA induces apoptosis of prostate cancer cells DU-145 and PC-3 via Akt/Fox03a pathwayMethods:Experiments were carried out with different concentrations of SAHA.The concentration of DU145 cells was l1,3,9?M.and the concentration of PC-3 cells was 0.5,2,8?M.The expression levels of cyclin A2,cyclin B,pAkt,pFox03a,Bim,Bcl-2,Bax and survivin in DU145 and PC-3 were detected by Western blot.The mRNA expression levels of Bim and Fas-L were detected by qRT-PCR.The expression of FoxO3a in DU145 and PC-3 cells was silenced by siRNA,and the expression levels of Fox03a,Bim,Bcl-2,Bax and survivin were analyzed by Western blot.RESULTS:In DU145 and PC-3 cells,the expression levels of cyclin B,cyclin A2,pAkt,pFox03a,Bcl-2 and survivin were significantly lower in SAHA than in the control group(P<0.05).The expression level of Bax was statistically significant compared with the control group(P<0.05).The results of qRT-PCR showed that the mRNA expression levels of Bim and Fas-L were significantly higher than those of the normal control group(P<0.05).In DU145 and PC-3 cells,.si-Fox03a successfully reduced the protein expression of Fox03a compared with the control group(P<0.05).After the decrease of Fox03a expression level,the effect of SAHA on Bcl-2 family protein and Survivin was observed.Partially eliminated.Compared with the control group,si-Fox03a partially restored the expression of Bcl-2 and Survivin,and decreased the expression levels of Bim and Bax.The ratio of Bcl-2/Bax increased after the activation of Fox03a and was statistically significant compared with the control group.(P<0.05).Part ? Effect of SAHA combined with docetaxel on apoptosis of prostate cancer cells DU-145 and PC-3METHODS:DU 145 cells were divided into three groups for subsequent experiments.The control group,docetaxel(DT)group,docetaxel(DT)+ SAHA(3 ?M)were used to separate PC-3 cells.The subsequent experiments were performed in three groups:control group,docetaxel(DT)group,docetaxel(DT)+ SAHA(2(?M);docetaxel concentration was 100 nmol/L,and the intervention time was After 48 hours,the effects of SAHA and docetaxel(DT)on the viability of prostate cancer cells were detected by MTT assay,and apoptosis was detected by Annexin-V-FITC/Pl double staining.RESULTS:The results of DU-145 cells showed that the cell viability of DT group was significantly lower than that of the control group(P<0.05).The cell viability of DT+SAHA group was significantly lower than that of DT+SAHA group.Statistical significance(P<0.05);PC-3 results showed that the apoptosis rate of DT group was significantly higher than that of the control group(P<0.05);DT+ was compared with DT+SAHA group.The apoptosis rate of SAHA group was significantly increased and the difference was statistically significant(P<0.05).Conclusion:1.SAHA inhibited the viability of DU-145 and PC-3 cells,and SAHA induced cell cycle arrest of DU-145 and PC-3 cells in G2/M phase in a dose-dependent manner;2.SAHA induces apoptosis of DU-145 and PC-3 cells;3.SAHA may induce apoptosis of prostate cancer cells through Akt/Fox03a pathway;4.SAHA can enhance the inhibition of proliferation of DU-145 and PC-3 cells by docetaxel and induce apoptosis of DU-145 and PC-3 cells. |
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