Luminex液相芯片技术测定氨甲喋呤对映体耐药A549细胞株中磷酸化酪氨酸激酶受体  

Expression of phosphorylated tyrosine kinase receptors in methotrexate enantiomers-resistant A549 cell lines by Luminex multiplex analysis

作　　者：许维东[1] 何晓东[2] 孙利[2] 李道静[2] 张永娟[2] 张白银[2] 沈佐君[2] 

机构地区：[1]金华市人民医院检验科,浙江金华321000 [2]安徽省立医院安徽省临床检验中心,合肥230001

出　　处：《临床检验杂志》2011年第8期564-566,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的探讨氨甲喋呤(MTX)对映体诱导的耐药A549细胞株中磷酸化酪氨酸激酶受体(RTKs)的表达差异及受体间的相关性。方法用MTX对映体浓度递增结合低剂量持续诱导肺腺癌A549细胞株,用Luminex液相芯片技术检测L-(+)-MTX/A549细胞、D-(-)-MTX/A549细胞和亲本A549细胞3组细胞中7种磷酸化RTKs[磷酸化表皮生长因子受体(p-EGFR)、磷酸化胰岛素样生长因子1受体(p-IGF-1R)、磷酸化肝细胞生长因子受体(p-HGFR)、磷酸化血小板源性生长因子受体(p-PDGFRβ)、磷酸化人类表皮生长因子受体2(p-HER-2)、磷酸化血管内皮生长因子受体2(p-VEGFR2)、磷酸化血管生成素受体(p-Tie-2)]的表达水平,分析它们之间的相关性。结果 D-(-)-MTX/A549组p-EGFR、p-HER-2的平均荧光强度(MFI)分别为26.91±1.81、16.56±1.15;L-(+)-MTX/A549组的MFI分别为54.13±2.75、16.94±1.44;亲本细胞组的MFI分别为54.38±2.88、33.75±2.49。p-EGFR与p-HER-2显著相关(P<0.05),p-HER-2和p-VEGFR、p-HGFR之间显著相关(P<0.05),其他磷酸化RTKs之间相关关系不显著。结论 MTX对映体诱导的耐药A549细胞株中,p-EGFR表达水平存在手性差异。Objective To study the differential expression of phosphorylated tyrosine kinase receptors（RTKs） and their correlation in MTX enantiomer-resistant cells.Methods A549 cells were exposed to intermittently and progressively increasing concentration of MTX enantiomers.The expressions of 7 phosphorylated RTKs（p-EGFR,p-IGF-1R,p-HGFR,p-PDGFRβ,p-HER-2,p-VEGFR2 and p-Tie-2） in the cells were detected by the Luminex xMAP system simultaneously,and their relationship was analyzed.Results The median fluorescent intensity（MFI） of p-EGFR and p-HER-2 were 26.91±1.81 and 16.56±1.15 in D-（-）-MTX/A549 cells,54.13±2.75 and 16.94±1.44 in L-（＋）-MTX/A549 cells,while 54.38±2.88 and 33.75±2.49 in parent cells.Significant correlation was noted between p-EGFR and p-HER-2（P0.05）.p-HER-2 was closely correlated with p-VEGFR and p-HGFR（P0.05）.The levels of other phosphorylated RTKs didn′t showed significant correlation in the cells of the three groups.Conclusion There was differential expression of p-EGFR in methotrexate enantiomer-resistant A549 cell lines.

关 键 词：氨甲蝶呤 A549细胞 对映体 酪氨酸激酶受体 液相芯片 

分 类 号：R73-37[医药卫生—肿瘤] R734.2[医药卫生—临床医学]