| Objective:To investigate the effect of dihydroartemisinin on expression of tumor suppressor gene UCHL1 in human prostate cancer PC-3 cell lines, and explore its regulation mechanism.Methods:PC-3 cells were treated with different concentrations (25,50, and 100μmol/L) of DHA for 48 h, while PC-3 cells without DHA treatment was used as the control group. Then the apoptosis and cell cycle distribution were detected by flow cytometry. The expressions and cellular locations of DNA methyltransferase 1 (DNMT1) and UCHL1 protein were detected by immunofluoresce-nce staining. The expression levels of UCHL1, DNMT1, phosphorylated Akt (p-Akt) and Akt proteins were detected by Western blotting; Wortmanin was used as a positive control group for comparing the expression of p-Akt protein in DHA group.Results:DHA induced the apoptosis of PC-3 cells, arrested the cell cycle at phase G2/M with significant differences compared with the control group (P< 0.05). After DHA treatment, DNMT1 protein translocated to the cytoplasms from the nuclei, and the overall expression level in the nuclei and cytoplasms was significantly decreased as compared with the control group (P<0.05); However UCHL1 protein level was significantly increased in cytoplasms of PC-3 cells treated with DHA (P< 0.05). Compared with the control group, UCHL1 expression was upregulated while DNMT1 expression was downregulated (both P< 0.05), and p-Akt expression level was reduced (P< 0.05), but Akt protein was not affected (P> 0.05) in the DHA treated group and positive control group. Downregulation of p-Akt expression was more significant in high concentration of DHA treatment group, more closer to that in the positive control group.Conclusion:DHA could inhibited DNMT1 expression, restored the function of tumor suppressor gene UCHL1, induce apoptosis and block cell cycle progression the mechanism may be related to the activity suppression of PI3K/Akt pathway. |
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