脑胶质瘤多药耐药机制的研究

Mechanism of glioma to multidrug resistance

为提高脑胶质瘤的化疗疗效 ,探讨胶质瘤的耐药机制 ,利用已建立的胶质瘤多药耐药细胞株 (C6 /adr) ,采用 RT- PCR法、免疫组织化学法分别对 C6及 C6 / adr细胞株 mdr- 1基因及其编码产物 P糖蛋白 (Pgp)进行了定性研究 ,同时采用流式细胞免疫学方法分别进行耐药蛋白及靶酶定量分析。结果显示 ,C6 mdr- 1基因表达阴性 ,C6 / adr mdr- 1基因表达阳性 ;C6 / adr Pgp强阳性 ,主要位于细胞膜及突起上。 C6、C6 / adr中的 Pgp分别为4.17%± 0 .6 3%、40 .5 9%± 4.77%(P<0 .0 1) ;其肺阻蛋白 (L RP)分别为 2 .92 %± 0 .2 2 %、2 1.17%± 1.79%(P<0 .0 1) ;拓扑异构酶 α(TOPO α)分别为 2 2 .88%± 1.94%、16 .77%± 1.0 8%(P>0 .0 5 ) ;谷胱甘肽 S-转移酶л(GST- л)分别为 5 .93%± 0 .78%、31.81%± 8.76 %(P<0 .0 1)。认为 Pgp、多药耐药相关蛋白 (MRP)、L RP在耐药胶质瘤中可共同表达 ,Pgp、MRP、L RP、GST-л在胶质瘤继发性耐药中起重要作用 ,测定其变化有助于化疗药物的选择。

To study the mechanism of glioma to multidrug resistance.C6 and C6/adr,mdr-1 gene of cell line and its expression Pgp were studied by RT-PCR,immunocytochemical technology,meanwhile C6 and C6/adr resistance protein(Pgp,MRP,LRP,GST-π,TOPOⅡα)were analyzed quantitatively by using flow cytmetric-immunologic method.Results showed C6/adr mdr-1 gene expressed positive,C6 mdr-1 gene expressed negative.The enlarged bond 157bp.Immunocytochemical technology revealed C6/adr Pgp positive strongly which was in the cell membrane.FCM Indicated C6 and C6/adr respectively(%):Pgp:4.17±0.63,40.59±4.77(P<0.01); MRP:3.59±0.13,14.14±1.01(P<0.01); LRP:2.92±0.22,21.17±1.79(P<0.01);TOPOⅡ:(22.88± 1.94,16.77±1.08(P>0.05); GST-π:5.93±0.78,31.81±8.76(P<0.01).These findings indicate that Pgp,MRP,LRP coexpress in multidrug resistance glioma.Pgp,MRP,LRP,GST-π may play important roles in acquired drug resistance of glioma and analysis of them can help to choose more effective chemotherapy drugs.