胃癌耐药细胞特异表达新基因的部分cDNA克隆

The partial cDNA cloning of a novel gene specifically expressed in drug-resistant human gastric cancer cells

目的:研究胃癌多药耐药(multipledrgresistance,MDR)相关基因。方法:应用差异显示技术,对人胃癌SGC7901细胞系及其长春新硷耐药细胞(SGC7901VCR)之间的差异表达基因进行了差异显示。结果:得到3个长春新硷耐药细胞特有的差异条带的cDNA克隆,命名为VRF1,VRF2与VRF3。RNA打点杂交证实,VRF1可在人胃癌SGC7901细胞中的长春新硷耐药细胞中特异表达,而在人胃癌非耐药的SGC7901细胞中不表达,可能与SGC7901肿瘤细胞耐药性产生有一定的关系。VRF2与VRF3为无差异的假阳性条带,本实验未对其做进一步地研究。结论:经序列测定分析,VRF1未在GenBank数据库中发现其同源序列,证实为一种新的基因序列。VRF1为3′端的非编码区序列,并有一个加尾信号AATAA。该序列已被GenBank数据库收录,编号为AI00266。

Aim : Toresearchrelatedgenesofdrug resistanthumangastriccancercellline .Methods:Differentiallyexpressed genesbetweenthehumangastriccancercelllineSGC 790 1andtheirvincristine resistantcells (SGC 790 1 VCR)havebeen differentiallydisplayedbydifferentialdisplaytechnique.Results:Threedifferentiallydisplayedbands(VRF1,VRF2and VRF3 )wereobtainedfromtheSGC 790 1 VCRCellsafterPAGE .Theywerere amplifiedandclonedintovectorpGEM 7Zf ( +)T A .RNAdotblothybridizationanalysisofthethreecDNAfragmentsshowedthattheVRF1wasspecificallyexpressed invincristine resistantSGC 790 1cells.Conclusion :SequencingdemonstratedthattheVRF1genefragmenthasnohomolo goussequencetoitinGenBank ,suggestingthatitwasthepartialcDNAsequencesinanovelgene .VRF1genewasconfirmed tobe 3′terminaluntranslatedregion( 3′UTR)by 3frametranslationanalysisandinadditionalsohadanacanonicalpolyadeny lationsequence (AATAA) .TheVRF1genewasemployedbyGenBankasaserialnumberAI0 0 0 2 66.