Mechanism of Sepia Ink Polypeptide-induced Apoptosis in DU-145 Prostate Cancer Cells
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Abstract:
In this study, the mechanism of apoptosis induced by sepia ink polypeptide (SHP) in DU-145 prostate cancer cells was explored. The effect of SHP on the proliferation of DU-145 cells was examined by the Cell Counting Kit-8 (CCK-8) assay. Typical morphological changes in DU-145 cells were observed with hematoxylin and eosin (HE) and acridine orange/ethidium bromide (AO/EB) staining. The early-stage apoptosis rate was measure using flow cytometry (FCM), and the changes in the expression of apoptosis-related genes (p53, Bcl-2, Bax, Caspase-3, and VEGF) were evaluated via western blotting. The results showed that SHP significantly inhibited the proliferation of DU-145 cells in a time-and dose-dependent manner. DU-145 cells developed morphological features of apoptosis after treated with SHP.FCM studies revealed that the early-stage apoptosis rate of DU-145 cells increased from 12.25% to 34.20% with increasing SHP concentration and duration of treatment. Western blotting results showed that after 24 h treatment with SHP, the expression of anti-apoptotic proteins Bcl-2 and VEGF decreased, while the expression of p53, Bax, and Caspase-3 increased. Collectively, these results suggest that SHP induced apoptosis in DU-145 cells. The mechanism might involve the decrease in Bcl-2/Bax expression ratio by activation of the tumor suppressor gene p53. Moreover, VEGF expression was down regulated, and apoptotic protease Caspase-3 was activated, thus triggering the apoptosis cascade reaction.
