氨甲蝶呤对映体耐药A549细胞的表皮生长因子受体基因表达及迁移能力  

Expression of epidermal growth factor receptor gene in methotrexate enantiomers-resistant A549cells and influence on cellular migration ability

作　　者：张白银[1] 何晓东[1] 孙余婕[1] 张永娟[1] 嵇金陵[1] 沈佐君[1] 

机构地区：[1]安徽医科大学附属省立医院临床检验中心,合肥230001

出　　处：《临床检验杂志》2012年第5期375-377,385,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金资助项目(30672011);安徽省自然科学基金资助项目(050430902);安徽省人才开发资金资助项目(2005Z040)

摘　　要：目的研究氨甲蝶呤(MTX)对映体耐药人非小细胞肺癌A549细胞的迁移能力以及表皮生长因子受体(EGFR)mRNA的表达。方法用细胞划痕试验检测L-(+)-MTX/A549细胞和D-(-)-MTX/A549细胞的迁移能力;双层软琼脂克隆试验检测L-(+)-MTX/A549细胞和D-(-)-MTX/A549细胞的克隆形成率并观察集落的形态;用RT-PCR检测亲本A549细胞、L-(+)-MTX/A549细胞和D-(-)-MTX/A549细胞中EGFR mRNA的表达。结果加入MTX 72 h后D-(-)-MTX/A549细胞的迁移能力(1 230.1±40.2)高于L-(+)-MTX/A549细胞(530.3±25.4);D-(-)-MTX/A549细胞、L-(+)-MTX/A549细胞和亲本A549细胞的克隆形成率(%)分别为(1.38±0.17)、(1.36±0.13)和(1.37±0.15),差异无统计学意义(P>0.05);亲本A549细胞、L-(+)-MTX/A549细胞均有EGFR mRNA表达,其光密度值(IDV)分别为(6 630±64)、(3 697±27),差异有统计学意义(t=103.42,P<0.01)。而D-(-)-MTX/A549细胞不表达EGFR。结论 D-(-)-MTX诱导的A549细胞的迁移能力大于L-(+)-MTX。EGFR基因表达具有手性差异。Objective To investigate the migration ability of methotrexate（MTX） enantiomers-resistant non-small cell lung cancer（NSCLC） cell line A549 and the expression of epidermal growth factor receptor（EGFR） in the cells.Methods The migration ability of L-（＋）-MTX/A549 and D-（-）-MTX/A549 cells were evaluated by cell scratch assay.The colony formation rates and the morphology of cell cluster of L-（＋）-MTX/A549 and D-（-）-MTX/A549 were determined by double-layer soft agar colony formation assay.The mRNA expression of EGFR in parental A549 cells,L-（＋）-MTX/A549 cells,D-（-）-MTX/A549 cells were detected by RT-PCR.Results The migration ability of D-（-）-MTX/A549 cells（1 230.1±40.2） was stronger than that of L-（＋）-MTX/A549 cells（530.3±25.4） at 72 h after adding MTT.The rate of colony formation in D-（-）-MTX/A549,L-（＋）-MTX/A549 and parental A549 cells was（1.38±0.17）,（1.36±0.13） and（1.37±0.15） respectively.There was no statistically significant difference among D-（-）-MTX/A549,L-（＋）-MTX/A549 and parent A549 cells（P〉0.05）.The mRNA expression of EGFR in parental A549 cells was（6 630±64）,and in L-（＋）-MTX/A549 cells was（3 697±27）,thus statistical difference between the two groups was found（t=103.42,P〈0.01）,but the EGFR mRNA in D-（-）-MTX/A549 cells barely expressed.Conclusion A549 cells induced by D-（-）-MTX showed greater migration ability than those induced by L-（＋）-MTX.The expression of EGFR gene presented significant difference in different chirality.

关 键 词：表皮生长因子受体 氨甲蝶呤 对映体 肿瘤转移 

分 类 号：R817.4[医药卫生—影像医学与核医学]