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Effects of Chrysin on TRAIL-induced Apoptosis in Human Hepatocellular Carcinoma Bel-7402 Cells

Author: QiuYanJun
Tutor: ZhuJianSi
School: Nanhua University
Course: Pathology and Pathophysiology
Keywords: Hepatic carcinoma ChR Tumor necrosis factor-related apoptosis-inducing ligand Apoptosis Sensitizing effect
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 20
Quote: 0
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Abstract


Whether the purpose of research chrysin (chrysin, ChR) by upregulating death receptor 5 (Death receptor 5, DR5) Sensitizing tumor necrosis factor-related apoptosis-inducing ligand (TNF-related apoptosis-inducing ligand, TRAIL) - induced human hepatoma Bel- 7402 cell apoptosis . Methods Cultured human hepatoma Bel-7402 cells . MTT assay was used to determine the cell activity . Propidium iodide (PI) staining by flow cytometry (FCM) quantitative analysis of the rate of apoptosis . DNA agarose gel electrophoresis confirmatory induced apoptosis . Western Blotting detection cell DR5 protein expression . Results detected by MTT assay results show that , the single use ChR or TRAIL processing 48 hours of human hepatoma Bel-7402 cell proliferation inhibition activity median inhibitory concentration (IC50 values ??) were 252μmol / L and 664 ng / mL , however , with 40μmol / L ChR pre - incubation for 30 minutes , then add TRAIL 48 hours of treatment , the IC50 value 91 ng / mL , the CI value of the combined medication effect is 0.497 . The PI staining FCM analysis showed that the single with 40μmol / L ChR or 100 ng / mL of TRAIL role in 48 hours, apoptosis rate ( hypodiploid DNA content cell percentage ) 4.92% ± 0.38% and 6.32% ± 0.39 % , respectively , However, after the pre - incubated for 30 minutes with 40μmol / L ChR , further added to 100 ng / mL of TRAIL for 48 hours, apoptosis rate was 31.3 % ± 2.50 % ; DR5/Fc (1μg/ml) to the sub - toxic concentration of the the ChR used in combination with TRAIL apoptosis of human hepatoma Bel-7402 rate decreased to 15.2% from 32.4% ± 0.47% ± 0.38 % ( P lt; 0.05 ) . DNA agarose gel electrophoresis , 40μmol / L ChR pre- incubated for 30 minutes , then add 100 ng / mL TRAIL for 48 hours , in the human hepatoma Bel-7402 cells , demonstrating the typical DNA ladder map . Western Blot analysis of concentration and time -dependent manner , ChR raised Bel-7402 cells DR5 expression ; DR5 chimeric protein effectively raised ChR the Bel-7402 cells DR5 expression . Conclusion 1 sub - toxic concentrations of ChR with enhanced TRAIL-induced apoptosis of human hepatoma Bel-7402 cells . 2. ChR sensitized TRAIL-induced cell apoptosis of human hepatoma Bel-7402 mechanism with its raised DR5 expression .

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