Th1/Th2相关细胞因子基因检测方法的建立

Establishment of detecting methods of Th1/Th2 cytokine gene

建立检测Ｔｈ１／Ｔｈ２相关细胞因子的方法。方法：设计６对用于ＩＬ－２、ＩＬ－４、ＩＬ－１０、ＩＬ－１３、ＩＦＮｒ和β－ａｃｔｉｎ基因扩增的引物，建立可用于Ｔｈ１／Ｔｈ２相关细胞因子检测的ＲＴ－ＰＣＲ技术，同时用切口平移法制备６种细胞因子的ｃＤＮＡ探针，建立ＲＮＡ斑点杂交技术。结果：用ＲＴ－ＰＣＲ和ＲＮＡ斑点杂交技术检测２０种肿瘤细胞株，发现１１／２０为Ｔｈ２型细胞因子表达，８／２０为Ｔｈ０型，１／２０未检测到细胞因子。结论：ＲＴ－ＰＣＲ技术和ＲＮＡ斑点杂交技术是检测Ｔｈ１／Ｔｈ２相关细胞因子的有效方法。

Objective: TO establish methods for sensitive, rapid, economical detection of Th1/Th2 Cytokine gene expression. Method: sixpairs of Primers specific for the IL-2, IL-4, IL-10, IL-13, IFNr and B-actin were designed. A Sensitive and rapid method for detection of Th1/Th2cytokine genes by RT-RCR was established. Meanwhile another method: the RNA hybridization with sir cDNA probes were established. Results:The RT-PCR reaction and RNA hybridization was used to test five kinds of Cytokine mRNAs expressed 20 tumor cell lines on mRNAs were expressed in eleven Cell lines;The mRNAs in eight cell lines. Conclusion:The preliminary results suggested that RT-PCR and RNA hybridizationnight be used to test Th1/Th2 cytokine gene expression.