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本研究探讨了BGFE过量表达对人鼻咽癌细胞株CNE-2Z有关生物学效应的影响.将EGFR基因c-erbB_1 cDNA5’端编码区1350 bp长的片段以3’→5’反向插入逆转录病毒载体(pLXSN),构建成反向重组质粒pLXSN/AS,通过PA317细胞得到高滴度达病毒并感染靶细胞CNE-2Z,同样经G418常规筛选,获得反义表达质粒转染的10个阳性克隆和空载体转染的3个阳性克隆,PCR证实外源基因和载体序列都能稳定地整合到受体细胞基因组内,对两个反义RNA表达的代表性细胞克隆CNE-2Z/AS(4)、CNE-2Z/AS(8)及空载体转染的阳性克隆CNE-2Z/pLXSN和对照细胞CNH-2Z分别进行分析.结果表明:在光镜下两个反义克隆细胞组细胞变大,胞内有空泡,且有的细胞呈现成纤维细胞形态;CNE-2Z/pLXSN与对照CNB-2Z相比,无明显差异.电镜下各实验组与对照组细胞形态没有明显差异.通过LIGAND和Scachard图分析,两个反义克隆细
This study was designed to investigate the effects of BGFE overexpression on the biological effects of CNE-2Z on human nasopharyngeal carcinoma cell line CNE-2Z.A gene fragment of 1350 bp encoding the 5 ’end of EGFR gene cDNA was reversely inserted in 3’ → 5 ’ The recombinant plasmid pLXSN / AS was constructed by reverse transcription polymerase chain reaction (pLXSN). The target virus CNE-2Z was infected by PA317 cells in high titer and infected with target cell line CNE-2Z by G418. Positive clones and empty vector transfected three positive clones, PCR confirmed that exogenous gene and vector sequence can be stably integrated into the recipient cell genome, the two representative antisense RNA expression cell clone CNE-2Z / AS (4), CNE-2Z / AS (8) and empty vector transfected CNE-2Z / pLXSN and control cells CNH-2Z were analyzed respectively.The results showed that under the light microscope two antisense clone cells CNE-2Z / pLXSN had no significant difference compared with that of control CNB-2Z. There was no significant difference in cell morphology between the experimental group and the control group under electron microscope. By LIGAND and Scachard graph analysis, two antisense clones