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| The Mechanisms of Panobinostat Reversing HepaCAM Gene Expression in Prostate Cancer |
| CHEN Er1, OU Li-ping1, TANG Min1, LIU Nan-jing1, WU Xiao-hou2, LUO Chun-li1 |
1. Key Laboratory of Clinical Laboratory Diagnostics of Ministry of Education, Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China;
2. Department of Urinary Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China |
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Abstract Objective: To study the reversing effect of Panobinostat on the expression of tumor suppressor gene hepatocyte cell adhesion molecule (hepaCAM) and its synergistic inhibitory effect with hepaCAM on the growth of prostate cancer (PCa) cells. Methods: PC3 cells cultured in vitro were treated with Panobinostat at different concentrations and the effect of Panobinostat on cell proliferation was detected by MTT assay, followed by the expression changes of hepaCAM, histone deacetylase (HDACs), and the acetylation of lys9 of histone H3 (Ac-H3K9) detected by RT-PCR and Western blot. Then after the cells were treated with different factors, cell proliferation activity was detected by MTT assay, cell cycle change was detected by flow cytometry, and gene expressions of cell cycle regulating factors CyclinD1 and proliferating cell nuclear antigen (PCNA) were detected by RT-PCR and Western blot. Results: The inhibitory effect of Panobinostat on the growth of PC3 cells was positively correlated to its concentration and duration. The expression levels of hepaCAM mRNA, its protein, and intranuclear Ac-H3K9 protein were increased, while that of both mRNA and protein of HDAC1, HDAC3, and HDAC4 were decreased when the concentration of Panobinostat increased; Cell growth was significantly inhibited in both the adenovirus-hepaCAM adenovirus alone group and the Panobinostat alone group, with an inhibition rate increased along with the prolonged action time. This inhibition became even particularly noticeable when the two were combined, with a difference of statistical significance (P < 0.05); the percentage of cells in S phase was significantly higher by the combined treatment compared with ad-hepaCAM alone and Panobinostat alone, respectively, with statistically significant differences (P < 0.05). This combination could further downregulate the mRNA and protein expressions of cyclinD1 and PCNA, and the difference was statistically significant (P < 0.05).Conclusion: Panobinostat may enhance the acetylation of lys9 of histone 3 and reverse the hepaCAM expression through its inhibitory effect on HDACs activity in PCa PC3 cells; hepaCAM-expressed adenovirus combined with Panobinostat may synergistically inhibit the growth of PC3 cells, via a potential mechanism associated with the down-regulation of the expression of CyclinD1 and PCNA. This reveals the reasons of hepaCAM deficiency in tumors and provides scientific support for the application of Panobinostat in clinical therapy.
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Received: 10 March 2016
Published: 25 June 2016
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