自发永生性转化人胚肺上皮细胞系HLEC的建立及鉴定

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1、第36卷第2期2005年4月解剖学报ACTAANATOMICASINICAVol.36,No.2Apr.2005自发永生性转化人胚肺上皮细胞系HLEC的建立及鉴定张志谦*胡颖王冰晶赵威林仲翔(北京大学临床肿他学院賢北京市肿制防治研究所细胞生物塔•北京100034)［摘要］目的对源自人胚肺组织原代培养、形态似上皮类型的细胞系一人肺上皮细胞(HLEC)的牛物学转性进行鉴定，为相关研究和应用提供模型和依据"方法分别采用相基显微镜下观察、细胞计数、卩•半乳糖昔酬染色、免疫荧光细胞化学。人Ahi序列PCR扩增、染色体计数、软晾脂集榕形成实验和裸鼠致瘤实验对HLEC的形态学待征、生

2、长特性、衰老状态、上皮细胞标志物、来源、细胞遗传学特点和恶性转化等生物学特性进行研究鉴定。结果HLEC角蛋白和上皮堕细胞间黏附分子钙黏着索E-cadherin免疫荧光染色阳性，染色体众数在41~44,人Ah序列PCR扩增结果阳性，p•半乳糖伴酶阳性细胞的比例未见晦细胞代数的增加而升离，软琼脂和裸鼠体内均不生长，微丝及黏着斑结构正常。结论HIEC细胞具有永生化细胞特征、属相对正常人肺上皮亚二倍体细胞，可用于肺癌癌变机理研究和癌细胞的相对正常对照。［关權词］人肺上皮细胞；不死性转化;钙黏着素；角蛋白；亚二倍体［中图分类号］Q249；R37-73［文献标识码］A［文章编号］0

3、529-1356(2005)02-113ESTABLISHMENTANDCHARACTERIZATIONOFASPONTANEOUSLYIMMORTALIZEDHUMANLUNGEPITHELIALCELLLINEHLECZHANGZhi-qian*,HUYing,WANGBing-jing,ZHAOWei,LINZhong-xiang(DepanmenlofCellBiology,theSchoolofOncology,PekingUniversity>BeijingIrutUuieforCancerResearchvBeijing100034,China)[Abst

4、ract]ObjectiveTheaimofthisstudywastocharacterizethebiologicalnaturesofhumanlungepithelialcells(HLEC),whichweresubculturedfromhumanfetallungprimaryculture・MethodsHIJECcellswereassayedfortheirgrowthcharactersbymeasuringpopulationdoublingtimeandsoft・agarcolonyformingexperiment・Epithelialmol

5、ecularmarkersweredetectedbykeratinandE-cadherinimmunofluorescentstaining・ChromosomemedalnumberandhumanAlu・sequencebasedPCRwerealsoperformedtocharacterizetheircytogencUcnatureandspecieorigin・Hiecellswerefurtherinoculatedintonudemicetoexcludetumorigenecity・Finally,p-galactosidasestainingwa

6、susedforevaluatingitssenescenceMatus.ResultsHLECcells)ia(iconstantpopulationdoublingtimeamongdifferentpassage趴andveryfewp-galactosidases(x)silivecelkwerefoundeveninculturesover100paBsages・Theywerepositiveforkeratin,E-cadherintwithnormalstructureofmicrofilaments・Chromosome(medalnumberwas4

7、1・44)andAlu-PCRverifieditshumanorigin.Furthermore,theywerenottumorigeneticinnudemice.ConclusionHImECcellsares

8、x)ntancouslyimmortali^d9sub-diploidhumanlungepithelialcells・’Hieycanbeusedasamodelfortheresearchoflungcancer・[Keywords

9、Humanlungepithelialcell;Immortalization;Ker