人骨肉瘤差异表达基因的筛选

目的 研究人骨肉瘤组织与正常骨组织之间的差异表达基因,探讨骨肉瘤的基因改变,为建立骨肉瘤的基因诊断方法打下基础。方法 采用mRNA差异展示技术,应用3条锚定引物和8条随机引物从人骨肉瘤组织与正常骨组织中分离出差异表达基因,对其进行,是序,用打点杂交技术证实其是否为差异表达基因,将筛选出差异表达基因在GenBank检索进行序列同源性分析。结果 筛选及克隆出6条差异条带,为ZOL03,ZOL51,1C82,1C74,2A21及2G12,RNA打点杂交证实它们在骨肉瘤中表达,在正常骨组织中不表达。6条差异片段经序列测定,在GenBank数据库中进行序列相似性分析,证实ZOL03,ZOL51同源性极低,1C82,1C74,2A21,2G12同源性较低,此6条片段为新基因序列ORF finder分析未发现具有开放读框,均属于非编码区序列,其中ZOL03和ZOL51两个序列在GenBank数据库中登录,接受号为BI894276和BI936370。结论 分离并克隆了人骨肉瘤组织与正常骨组织之间的6条差异表达基因,同源性差,均确认为新基因,可能是骨肉瘤差异表达相关基因。

Screening of differentially expressed genes in human osteosarcoma tissue

AIM To explore differentially expressed genes between human osteosarcoma tissue and normal bone tissue and to discuss the alteration of genes in osteosarcoma. METHODS By way of mRNA differential display PCR, using 3 anchor primers and 8 stochastic primers, differentially expressed genes between human osteosarcoma tissue and normal bone tissue were isolated, cloned and sequenced. Then the genes were approved with technique of dotblot, searched and analyzed against GenBank. RESULTS Six genes were screened and cloned. RNA dotblot approved that they were expressed in osteosarcoma tissue and non expressed or lowly expressed in normal bone tissue. Sequencing and searching in GenBank for comparability found that the 6 sequences were partially similar to sequences in GenBank. 6 sequences were new genes and 2 of them were accepted by GenBank, the accepted numbers were BI894276 and BI936370. CONCLUSION 6 new differentially expressed genes between human osteosarcoma and normal bone tissues were isolated and cloned with low comparability. They are probably correlative differential expressed genes of human osteosarcoma.