膀胱癌组织某一特异性EST片段全长扩增及生物信息学分析

Amplification and bioinformatics analysis of a specific EST fragment of bladder cancer tissues

目的对膀胱癌组织某一特异性表达序列标签(EST)片段进行全长扩增及生物信息学分析。方法收集1例进行手术的膀胱癌患者(病理证实为膀胱移行细胞癌)的癌组织,从新鲜的膀胱癌组织中提取RNA,利用RACE方法获取前期研究发现的某一特异性EST片段5'端和3'端DNA序列,通过Gene Tool软件合成RACE测序结果。对全长序列及其相应蛋白进行生物信息学分析,利用sequin软件编辑分析结果,并向NCBI提交获取基因号及蛋白号。结果成功获得该特异性EST片段的5'端和3'端,利用Gene Tool软件合成一个1 839 bp的全长序列,初步分析其含有完整的开放阅读框(ORF),染色体定位于17p13.3。该序列在Gene Bank获得基因号为KR780251,ORF翻译的多肽序列获得蛋白ID为AKO62675。结论该膀胱癌组织特异性EST片段基因号为KR780251,全长序列为1 839 bp,含有完整的ORF,其编码蛋白AKO62675可能是正常膀胱组织的基因突变体。

Objective To amplify the total length of a specific expressed sequence tag(EST)fragment of bladder cancer and to perform the bioinformatics analysis on it.Methods We collected 1 case of bladder cancer tissues which were pathologically confirmed bladder transitional cell carcinoma.The RNA from the bladder cancer tissues was extracted,and then we obtained the 5′end DNA sequence and 3’end DNA sequence of the EST fragment by RACE,and used the GeneTool to synthesize the results of DNA sequence.We analyzed the full length sequence of cDNA and its corresponding protein with bioinformatics,and submitted it to the NCBI to get GenBank accession number and protein ID by sequin software.Results We successfully got the 5′end and 3′end of the specific EST fragment,and used the GeneTool to synthesize a full sequence of 1839bp,and it had integrated ORF,chromosome was located in 17p13.3.We got the GenBank accession number,that was KR780251 in NCBI,and the ID of polypeptide sequence acquired protein translationed by ORF was AKO62675.Conclusion The gene of bladder cancer tissue-specific EST fragment is KR780251 with a full-length sequence of 1839 bp and contains an intact ORF.The encoded protein AKO62675 may be a mutant from the normal bladder tissues.