人膀胱癌相关基因cDNA片段的克隆与鉴定

目的　分离和鉴定与人原发性膀胱移行细胞癌发生发展相关的新基因。　方法　采用mRNA差异显示技术 ,以 8例原发性膀胱移行细胞癌组织及自身远离肿瘤的正常膀胱粘膜上皮为对照 ,通过基因表达的比较 ,找出差异条带进行再扩增、克隆及DNA序列分析 ,并通过Genbank数据库进行同源性检索。　结果　 (1)共分离并回收差异条带 37条。其中正常组织不表达或低表达而肿瘤组织出现表达或表达增强者 36条 ,正常组织表达而肿瘤组织失表达者 1条。 (2 )差异条带中与TBX3基因 (T box3基因 )同源 1条 ,同源性 99% ;与铜 ,锌 超氧化物歧化酶基因 (Cu ,Zn SOD基因 )同源 1条 ,同源性 99% ;与Ca2 激活氯离子通道基因家族成员 4 (cl.Ca4基因 )同源 1条 ,同源性 99% ;3条与人类已知基因或片段无明显同源 ,分别命名为YHHQ1,XHL ,LHX。　结论　Cu ,Zn SOD基因可能是新的与膀胱癌相关的癌基因。TBX3及cl.Ca4基因可能与人膀胱癌有相关性。YHHQ1可能是膀胱癌相关的候选抑癌基因 ;XHL ,LHX可能是与膀胱癌相关的候选癌基因

The clone and identification of cDNA fragments of relevant gene of human bladder cancer

Objective To isolate and identificate a specific new gene highly related to human bladder cancer and to investigate the carcinogenesis mechanism and genetic susceptibility. Methods Normal bladder epithelia and cancer tissue collected from 8 cases of primary bladder transitional cell carcinoma were studied using the high performance?sensitive technique of mRNA differential display.The differential fragments were found by comparing with genetic expression. 6 of them were reamplificated?cloned?sequenced,and performed the autoploidy index in the public database of genbank. Results (1)37 differential fragments have been isolated.36 of which are expressed or highly expressed in cancer tissues,but not so or lowly expressed in normal tissues;while it has been expressed in 1 normal tissue but not in the cancer tissue.(2)Results of indexing:1 is homologous to TBX3 (T box gene),autoploidy 99%.1 is homologous to cl.Ca4 gene (member 4 of Calcium activated chloride channel gene family), autoploidy 99%.1 is homologous to Cu,Zn SOD gene (Cu,Zn superoxide dismutase),99%.The remaining fragments are not homologous to unknown sequences in the public database of genbank,assigned as YHHQ1,XHL,LHX. Conclusions (1)It is verified at genetic level the relation between the conment's increasing of Cu,Zn SOD in bladder cancer derived tissue and the occurrence,advance of bladder cancer.(2)The relation between TBX3,cl.Ca4 gene and human carcinoma has been observed.(3)YHHQ1 may be the new candidate of anti oncogene related to bladder cancer.XHL and LHX may be the new candidates of oncogene related to bladder cancer.