红芪多糖对S180细胞体内化疗增效作用的差异蛋白质研究

目的:研究红芪多糖对S180瘤细胞化疗协同增效作用的差异蛋白质。方法:以环磷酰胺和红芪多糖联合环磷酰胺治疗S180荷瘤小鼠14天后测定小鼠免疫器官指数及抑瘤率,利用双向电泳分离提取的瘤细胞总蛋白,考染显色后应用PDQuest8.0软件分析电泳图谱,找出差异表达的蛋白质。结果:与环磷酰胺(20 mg/kg)组相比,红芪多糖(200 mg/kg)联合环磷酰胺(20 mg/kg)可明显抑制小鼠S180瘤生长(P<0.05),显著减轻环磷酰胺所致免疫器官指数的下降(P<0.01);建立了背景清晰、重复性好、分辨率高的S180瘤细胞双向电泳图谱,环磷酰胺组与红芪多糖联合环磷酰胺组蛋白质点数分别为:776±22和721±16;平均匹配率分别为:91.13%和89.37%。比较分析环磷酰胺组与红芪多糖联合环磷酰胺组双向电泳图谱,筛选出差异明显的蛋白质点28个,其中8个表达上调,19个表达下调,1个仅在环磷酰胺组表达。结论:红芪多糖联合环磷酰胺可减轻其对S180荷瘤小鼠的免疫抑制作用,增强环磷酰胺对S180瘤细胞的化疗效果,其作用可能与筛选出的28个差异蛋白质点有关,对这些差异蛋白质点的鉴定将为揭示红芪多糖的化疗协同增效作用提供实验依据。

The study on differentially expressed protein of the synergy effect of Radix Hedysari polysaccharides combined with chemotherapy on S180 tumour cells in vivo

Objective: To study differentially expressed proteins of the synergy effect of Radix Hedysari polysaccharides(HPS) combined with cyclophsphamide(Cy) on s180 tumour cells.Methods: The S180-bearing mice were injected with Cy and HPS combined with Cy respectively for fourteen days,then the index of immunity organs and tumor inhibition rate were observed.The total proteins extracted from the Cy and HPS combined with Cy treated S180 cells in tumour-bearing mice were separated by means of immobilized pH gradient based on two dimentional gel electrophoresis(2-DE).After Coomassie brilliant blue G250 staining,2-DE image analysis was performed to study the differentially expressed proteins using PDQuest8.0 software.Results: Compared with Cy group,the HPS combined with Cy group showed obviously suppressed effect on the growth of S180 tumor(P < 0.05) and significantly reduced the decrease of immunity organs index(P < 0.01),caused by Cy.The clearer,reproducible,high resolution 2-DE images were established and obtained,776 ± 22 和 721 ± 16 protein spots were resolved respectively in the Cy and the HPS combined with Cy treated groups.The match rate were 91.13% and 89.37% respectively in the Cy and HPS combined with Cy treated groups and twenty-eight differentially expressed protein spots were displayed between the Cy and HPS combined with Cy treated groups,of which eight showed higher expression,nineteen decreased in abundance,an only one disappeared in the HPS combined with Cy treated group.Conclusion: The HPS combined with Cy reduced immunosuppression caused by Cy on S180bearing mice and strengthened chemotherapy effect of Cy on S180 tumour cells,which might be related to the twenty-eight differentially expressed protein spots.Further identification of the differentially expressed protein spots may provide clues for the molecular mechanism of synergy effect of HPS Combined with Cy.