Study Of Effects And Mechanisms Of Panhenolide On Prolifertion And Apoptosis In Human Prostate Cancer DU-145Cells In Vitro

Objective:In recent years, the incidence and mortality of prostate cancer in China is also on the rise. The early days of the disease can be treated surgically. Because tumor development depends on the presence of androgen,Missed the times of surgery the majority of patients feasible androgen deprivation therapy, However, the time of treatment tumor often transformed into hormone-resistant prostate cancer(HRPC). Prostate cancer is not sensitive to the chemotherapy, and long ago lost the chance of operation. Therefore continued to develop an effective control of tumor growth and to improve the patient’s symptoms, drug having significance. Parthenolide (PTL) is a sesquiterpene dilute lactones, extracted from Chinese herbs tansy early years in some Western countries, has been purified and used for the anti-inflammatory analgesic. Is generally used to treat arthritis, headache, migraine, cold and fever, it has now been confirmed with the in-depth study of the domestic and foreign scholars feverfew, parthenolide has a significant inhibitory effect on a variety of tumor cells in vivo, such as breast cancer gastric cancer and liver cancer and other tumors. In recent years, some foreign studies show that the PTL may a role in prostate cancer, but few studies and floating on the surface, the specific role of the size and the mechanism is unclear, therefore this experiment by observing the PTL on cultured human androgen-independent prostate cancer DU-145cell proliferation, apoptosis, and to explore its mechanism, designed to provide experimental evidence for the adjuvant treatment of prostate cancer and the clinical applications of the Methods:1Cell culture MTT and cell count analysis of PTL can significantly inhibit the proliferation of DU-145cells, a time-and concentration-dependent manner, was a significant positive correlation (P<0.05). The immunocytochemistry staining results showed that, PTL treatment of DU-145cell proliferation marker K167lower protein concentration a negative correlation (P<0.05).2Proliferation analysis Respectively MTT assay and cell counting method to analyze the cell proliferation. Absorbance OD values of cells to determine the detected concentration, the time point, calculate the inhibition rate, Rate PTL affect the DU-145cell proliferation.3Flow cytometry analysis Take sensitive concentration, concentration gradient packet processing cells in each group after digestion prepared into a single cell suspension, and then after Having fixed, washing and dyeing process with flow cytometric analysis of the PTL DU-145cell cycle distribution and witheredthe impact of the death.4Immunocytochemistry staining The cells were seeded on glass slides treated with PTL48h, remove slides the cells in each group is fixed. Slides were treated with Triton X-100, closed after the first antibody and second antibody binding reaction, and then through the DAB color light microscope, count the number of positive cells, and to calculate the proportion of detection of KI67expression rates.5Western blotting Take the same amount of total cellular protein, pipette SDS-polyacrylamide gel electrophoresis electricity transfer film closed the primary antibody and secondary antibody, chemiluminescence process, Cleaved-caspase3、Bcl-2and Bax protein detection. Results:1PTL inhibits DU-145cell proliferation MTT and cell count analysis of PTL can significantly inhibit the proliferation of DU-145cells, a time-and concentration-dependent manner, was a significant positive correlation (P<0.05). The immunocytochemistry staining results showed that, PTL treatment of DU-145cell proliferation marker KI67lower protein concentration a negative correlation (P<0.05).2The PTL to promote apoptosis in DU-145The results of flow cytometry, The PTL to promote DU-145cells were arrested in G0/G1phase (P<0.01), and DU-145cells induced obvious apoptosis peak (P<0.05). Western blot analysis showed that PTL treatment of DU-145cell proliferation and apoptosis marker Cleaved-caspase3expression increases with the concentration of positive correlation (P<0.01). Western blot analysis showed that the process DU-145cells in PTL promote apoptotic protein Bax expression level was significantly higher, and concentration into a positive correlation (P<0.05), while the anti-apoptotic protein Bcl-2expression level was significantly lower (P<0.01).Conclusions:1Parthenolide significantly inhibited human prostate cancer DU-145cell proliferation, and showed the dose-dependent and may be lowered KI67protein.2Parthenolide on human prostate cancer DU-145cells, cell cycle. With the increase of the concentration of the drug big or small, DU-145cells in G0/G1phase of the cell number increased significantly decrease in S and G2/M phase cell number, cell cycle arrest in G0/G1phase.3Parthenolide can iduce apoptosis in prostate cancer DU-145, in a time-and dose-dependent manner. Aponptosis Cleaved-caspase3expression in prostate cancer DU-145cells may be related to the upregulation people. 4Regulation of Bax and downregulation of Bcl-2expression levels may be one of the mechanisms of Parthenolide-induced apoptosis in DU-145cells.