摘要
目的 :建立高表达 β 葡萄糖醛酸苷酶 (以下简称 βG)基因的肾癌细胞模型 ,并观察转染细胞的生物学特性。方法 :构建真核表达载体 pcDNA3.1 βG ,并利用阳离子脂质体介导将其转入人肾癌细胞株GRC 1中。用mRNA打点杂交和Westernblot,鉴定其转录与表达水平 ,并通过光镜、电镜及细胞增殖周期检测等方法 ,观察转染细胞的生物学特性。结果 :在mRNA与蛋白水平证实 ,转染细胞内有 βG基因的高表达。透射电镜观察 ,转染细胞中的溶酶体、内质网丰富 ,细胞微绒毛及突起增多 ,但转染前后肾癌细胞GRC 1的周期及生长情况无显著性差别。结论 :应用脂质体介导法 ,将βG基因导入人肾癌GRC 1细胞后 ,获得生物学特性稳定 βG基因高表达的肾癌细胞模型 。
AIM :Toestablisharenalcarcinomacelllinewhichcanhighly expressβ glucuronidase(βG) ,andtoobservethebiological characteristicsofthetransfectedcells .METHODS :Recombi nanteukaryoticexpressionvectorpcDNA3.1 βGwasconstruct ed .ItwastransfectedintorenalcancercellsGRC 1vialipo some .ThetranscriptionandexpressionofβGgeneweredetect edbydotblotandWesternblot.Thebiologicalcharacteristics oftheβGgene transfectedcellswasobservedunderlightmi croscope ,transmissionelectronmicroscopeandflowcytome try .RESULTS :DotblotandWesternblotdetectionconfirmed thattheβG genehadbeenstablyintegratedintothegenomic DNAoftheGRC 1cellsandwashighlyexpressed .Transmis sionelectronmicroscopeobservationshowedthatthelysosomes andendoplasmicreticulumwereabundant,thenumberofmi croviliandprocesswassignificantlyincreasedinthetransfected cells,butgrowthconditionandcellcycleofGRC 1cellshadno notabledifferencebeforeandaftertransfection. CONCLUSION :Arenalcarcinomacelllinethatcanhighlyex pressβG genewasestablished ,whichlaysthefoundationfor furtherstudyongenetherapy .
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2003年第2期145-147,共3页
Chinese Journal of Cellular and Molecular Immunology
