简介：Objective:Toclonemultidrugresistance(MDR)relatedgenesinlungadenocarcinomacelllines.Methods:ThedifferentiallyexpressedcDNAfragmentsbetweenA549andA549DDPcellswereanalyzedbymRNAdifferentialdisplayPCR(DDRT-PCR).ThefragmentsthusobtainedwerefurtheranalyzedbyDNAsequencingandNorthernblotting.Results:ThreedifferentiallyexpressedcDNAfragmentswereobtainedandconfirmedbyNorthernblot.Sequenceanalysisrevealedthattwoofthemwerenovelandonewas100%identicalwithICEgene.Conclusion:AnalyzingdifferentiallyexpressedfragmentbetweenA549andA549DDPcellsmaybehelpfulforfindingnewMDRrelatedgenes.ThedrugresistanceofA549DDPcellsmayberelatedtotheinhibitionordown-regulationofICEgene.

简介：目的：克隆COPS3基因cDNA,构建其原核融合蛋白表达载体并表达和鉴定。方法：从培养的人骨肉瘤细胞系SOSP-9607细胞中提取总RNA,经RT-PCR获得COPS3基因。将该基因克隆到pGEM-T-Easy载体中,酶切及测序鉴定。将COPS3基因插入pET28a融合蛋白表达载体中,IPTG诱导表达,进行SDS-PAGE分析。免疫印迹法鉴定COPS3蛋白的表达。结果：cDNA测序证明,获得了COPS3基因cDNA,其序列与Genebank中报道序列完全一致。酶切分析表明,成功构建了含COPS3基因的pET28a融合蛋白表达载体。SDS-PAGE以及免疫印迹法鉴定分析表明,COPS3蛋白获得高效表达,分子质量为51Ku,表达量约占菌体总蛋白的30%。结论：成功克隆和表达了COPS3cDNA。

简介：Objective:Toanalyzethechangesofgeneexpressioninphenylbutyrateinduceddifferentiationofgliomacells.Methods:Theexpressionlevelsof14000genesingliomacellsbeforeandafterinducementwithsodiumphenyl-butyratefor2hor6dayswereevaluatedbycDNAarraytechniqueandprovedbymulti-dotblotting.Results:expressionof98genesingliomacellsshowedchangesaftertheinducement.Somegenesinvolvedintranscriptionandtranslationandsomeoncogenesaredown-regulated,whilesomegeneinvolvedindifferentiationorapoptosisareup-regulated.18unknownexpressionsequencingtag(EST)changedtoo.Conclusion:Ageneexpressionprofileassociatedwithdifferentiationofgliomacellswasestablished.

简介：Asyntheticpolypeptide,pt27,whichisencodedbyacDNAclonewithantloncogeneactivity,p14-6,isfoundtobeabletoreduceremarkablythesoftagarcolonyformationabilityofpartofDTcellsandtoraisetheirresistancetotheouabalntoxtcity.Thisshowsthatthept27peptidecanaffecttheDTcellsInamannersimilartothep14-6doneandprovidesevidencethattherevertingactionofthep14-6toDTcellsmaybeexertedbytheexpressionofitscDNA.

简介：Objective:ToanalyzethedifferentiallyexpressedcDNAsequencesrelatedtochlorophyllin(CHL)mediatedinhibitionofmalignanttransformationofhumanbronchia1epithelialcellline(16HBE).Methods:16HBEcellstreatedwithchlorophyllinandanti-BPDEwereconductedastester,16HBEcellstreatedonlywithanti-BPDEwereconductedasdriver,andcDNArepresentationaldifferenceanalysis(cDNARDA)wasusedtocomparethedifferentialgeneexpressionbetweenthetwokindsofcells.ThecDNAfragmentswereligatedtopGEM-TvectorandtransformedintoJM109bacteria.TheplasmidDNAwassequencedandcomparedwithdatabaseinGenBankbyBLASTN.Results:Amongthe5clonedcDNAsequences,threewerenovelandwereregisteredindbESTdatabase,twoshowedsequencehomologytoalpha-enolaseandanewlyfoundgeneribosomalproteinS18/S6-like.Conclusion:These5cDNAsequencesmightplayimportantrolesinantitransformingeffectofchlorophyllin.

简介：目的探讨DNA修复基因RAD523＇非翻译区（3＇-untranslatedregion,3＇-UTR）miRNA靶序列单核苷酸多态性（singlenucleotidepolymorphism,SNP）与广西地区人群肝细胞癌（hepatocellularcarcinoma,HCC）遗传易感性的关系。方法采用病例-对照研究,对1002例确诊的HCC新发病例和1013例非肿瘤患者RAD52基因3＇-UTR区域miRNA靶序列SNPs（rs1051669、rs1051672、rs7301931和rs7310449）进行基因分型,并分析其基因型频率分布及其与HCC遗传易感性的关系。结果RAD52基因各SNP基因型在病例组和对照组中的分布频率差异均无统计学意义（P〉0.05）。调整年龄、性别、吸烟、饮酒和HBV感染等因素后,未发现各SNP与HCC易感性有关联;分层分析发现,在女性人群中,与携带rs1051669C等位基因相比,TT基因型可显著降低个体罹患HCC的风险（TTvsCT/CC：OR=0.03,95%CI：0.00～0.62,P=0.03）;与携带rs1051672G等位基因相比,AA基因型可显著降低个体罹患HCC的风险（AAvsGA/GG：OR=0.03,95%CI：0.01～0.88,P=0.04）。结论RAD52基因3＇-UTR区域miRNA靶序列SNPsrs1051669、rs1051672位点可能与广西地区女性人群HCC易感性有关。