摘要
目的研究红枣多糖对体外培养肝癌细胞增殖的抑制作用并初步探究其可能的作用机理。方法采用MTT法测定红枣多糖对体外培养的人肝癌细胞HepG2增殖的抑制作用;流式细胞术检测红枣多糖对人肝癌细胞HepG2周期和凋亡的影响;Real time RT-PCR检测红枣多糖对人肝癌细胞HepG2中Bcl-2和caspase3mRNA表达的影响。结果 MTT检测发现随着药物浓度的增高OD值呈现梯度递减,红枣多糖对HepG2的IC50=13mg/mL,最高浓度40mg/mL下所得最大抑制率为68.79%;流式细胞仪检测细胞凋亡结果可见早期凋亡率随药物浓度的增加而变大;流式细胞周期分析结果可见G0-G1期细胞数逐渐增多,S期细胞数有下降趋势,并有剂量依赖性;Real time RT-PCR检测发现Bcl-2凋亡抑制基因mRNA表达随药物浓度增高而降低,而凋亡关键基因caspase-3mRNA的表达随药物浓度增高而升高。结论红枣多糖对体外培养的肝癌细胞增值具有抑制作用,将肝癌细胞HepG2阻滞于G1期,并通过下调Bc 1-2而上调caspase-3mRNA表达诱导HepG2细胞凋亡。
Objective To study the multiplication effect of red dates polysaccharide on tumor cells in vitro and preliminarily explored its possible mechanism .Methods MTT method was used to determi-nate the value-added effect on human hepatoma cells HepG2 in vitro .Used the flow cytometric to de-tect the effect on human hepatocellular carcinoma cell cycle and apoptosis of red dates polysaccharide in HepG2 .Caspase3 and Bcl-2 mRNA were detected the effect on the expression in HepG2 cells by Real time RT-PCR .Results With the increased concentration of drug ,the OD decreased by the detection of MTT .The inhibition rate of IC50 （13 mg/mL） was 71 .52% .The detection of flow cytometric on ap-optotic cells showed early apoptosis rate increased as the concentration increased .From the result of cell cycle analysis ,the cell number gradually increased between time G0 and G1 ,and decreased in time S as the concentration of red datas polysaccharide was increased .The expression of apoptosis inhibi-ting gene Bcl-2 mRNA decreased when the concentration of drug increased from the analysis by Real Time RT-PCR .Conclusion The red dates polysaccharide has inhibition on cultured hepatoma cells and arrests the hepatoma cells HepG2 in phase G1 ,induces the apoptosis of HepG2 ,inhibits the expression of the mRNA Bcl-2 .
出处
《贵州医药》
CAS
2014年第6期506-508,共3页
Guizhou Medical Journal
