8-Br-cAMP和槲皮素联合应用对Eca-109细胞增变基因表型的影响  

Effects of 8-Br-cAMP ar quercetin combined use on changes of mutator phenotypes in Eca-109 cells

作　　者：宫璀璀[1] 郑乃刚[2] 王广兰[1] 吴景兰[2] 李璇[1] 

机构地区：[1]解放军第153医院检验中心,河南郑州450042 [2]郑州大学分子细胞生物学研究中心,河南郑州450052

出　　处：《河南医学研究》2010年第3期281-284,共4页Henan Medical Research

摘　　要：目的:探讨8-Br-cAMP和槲皮素联合应用对Eca-109细胞增变基因表型变动的影响。方法:将Eca-109细胞分为4组:①8-Br-cAMP组(Br组);②槲皮素组(Q组);③(Br+Q)组;④不加药物的对照组(C)。同时培养48 h,对以上4组细胞制备细胞滴片及硝酸纤维素膜(NCM)滴膜2种标本,分别进行Bcl-2、Bax、XRCC1、MRP和POLB的免疫组化和免疫斑点印迹实验,并以TUNEL法检测各组细胞凋亡率。结果:细胞凋亡率,C组为4%,(Br+Q)组为70%,Br组为42%,Q组为35%,(Br+Q)组>Br组或Q组>C组,(P<0.001)。Br、Q及(Br+Q)组与对照组相比,Bcl-2-IR及XRCC1-IR皆低于C组(P<0.001),而Bax-IR则高于C组(P<0.001);Br组:Q组或Br组:(Br+Q)组(P>0.05)。Bcl-2-IR与XRCC1-IR呈显著正相关,与Bax-IR和Bax-IR与XRCC1皆呈显著负相关。C组的MRP-IR、POLB-IR强于Br组、Q组或(Br+Q)组(P<0.01。)TLC扫描OD值:C组MRP-IR高于Br、Q或(Br+Q)组约3倍;POLB-IR高于Br、Q或(Br+Q)组约2倍;MRP与POLB呈显著正相关r=0.9838(P<0.01)。结论:Br与Q联合用药或单独用药皆可下调Bcl-2,XRCC1,MRP与POLB的表达,上调Bax的表达,提示Br与Q联合用药与单独用药皆可下调Eca-109细胞的增变基因表型。Objective： To study the effects of 8-Br-cAMP and quercetin alone or combined use on changes of mutator phenotypes in Eca-109 cells.Methods： The cultured Eca-109 cells randomized into 4 groups：8-Br-cAMP（Br） group,quercetin（Q） group,（Br＋Q） group,and Control（C） group treated with no any drugs.After cultured 48 h,two kinds of specimens including cell slide and NCM specimens were prepared and Bcl-2-IR,Bax-IR,XRCC1-IR,MRP and POLB were detected by immunohistochemistry and the immunodot blot technique.Besides,the cell apoptosis was detected with TUNEL method in each group.Results： The cell apoptotic percentage was 70% in（Br＋Q） group,42 % in Br group,35 % in Q group,and 4 % in C group.（Br＋Q）〉Br or Q〉C（P〈0.001）.When compared with C group,Bcl-2-IR and XRCC1-IR were decreased and Bax-IR increased in the other 3 groups（P〈0.001）,Br vs Q or Br vs（Q＋Br）（P〈0.05）.There waspositive correlation between Bcl-2-IR and XRCC1-IR,but negative correlated with Bax-IR（P〈0.01）;The immunoreactivity（IR） showed：MRP-IR,C〉Br,Q or （Br＋Q） group（P〈0.01）;POLB-IR,C〉Br,Q or（Br＋Q） group（P〈0.01）.There was a signify cant positive correlation between MRP and POLB.Conclusion： The Br combined with Q or single Br or Q lone could down regulate the expression of Bcl-2,XRCC1,MRP and POLB and up-regulate that of Bax,（Br＋Q） with highest cell apoptosis,which exhibites reduction of the mutator phenotype.

关 键 词：增变基因表型 8-BR-CAMP 槲皮素 ECA-109细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]