虎眼万年青皂苷类提取物对肝癌细胞miRNA的表达谱的影响

Analysis of expression of miRNA spectrum in hepatoma cells affected by saponins extract of Ornithogalum saudersiae

目的分析虎眼万年青皂苷类提取物(OSW-1)对肝癌细胞miRNA的表达谱的影响。方法细胞株分为四组:A组,Hep3B单克隆细胞系,作为对照组; B组,用200ng/ml OSW-1治疗单克隆细胞系24h; C组,用5000ng/ml多柔吡星治疗单克隆细胞系24h单克隆细胞系; D组,用80ng/ml OSW-1、2000ng/ml多柔吡星治疗24h的单克隆细胞系。使用第六代miRCURYTMLNA序列,Axon Gene Pix 4000B microarray scanner进行图像捕获,并使用折叠过滤鉴定差异表达的miRNA。结果 B组与A组比较,14种miRNA上调2倍以上,其中miR-125-1-3p,miR-299-5p及miR-1908上调4倍以上; 19种miRNA下调2倍以上,其中miR-208a和miR-126-3p下调30倍以上。D组与C组比较,25种miRNA上调2倍以上,其中miR-1275,miR-200c-3p和miR-142-3p上调20倍以上,尤其是miR-142-3p上调水平达到了58倍; 7种miRNA下调2倍以上,其中miR-3679-3p下调达到了10倍以上。结论 OSW-1影响众多miRNA表达,OSW-1协同抗肿瘤化疗药物,影响肝癌细胞生物学行为。

Objective To investigate the effect of the extract of saponins( OSW-1) on the expression of miRNA in human hepatoma cell. Methods The cell lines were divided into four groups: group A,Hep3 B monoclonal cell line,as control group;Group B was treated with 200 ng/ml OSW-1 for 24 hours;Group C,treated with 5000 ng/ml doxorubicin for 24 hours;Group D was treated with 80 ng/ml OSW-1 and 2000 ng/ml doxorubicin for 24 hours. Using the sixth generation miRCURYTMLNA sequence,Axon Gene Pix4000 B microarray scanner for image capture. The differentially expressed miRNA was identified by folding filter.? Results Compared with group A,14 miRNAs were more than 2 times up-regulated in group B,in which miR-125-1-3 p,miR-299-5 p and miR-1908 were up-regulated by more than 4 times;19 miRNAs were down-regulated by more than 2 times,in which miR-208 a and miR-126-3 p were down-regulated by more than 30 times. Compared with group C,25 miRNAs were more than 2 times up-regulated in group D,in which miR-1275,miR-200 c-3 p and miR-142-3 p were up-regulated by more than 20 times. In particular,the upregulation level of miR-142-3 p reached 58 times. 7 miRNA down-regulated more than 2 times,in which miR-3679-3 p down-regulated more than 10 times. Conclusion OSW-1 can affect the expression of miRNA,and OSW-1 in combination with anti-tumor chemotherapeutic drugs,affect the biological behavior of hepatoma cells.