吉非替尼对肺腺癌细胞增殖和表皮生长因子受体表达的影响  

The effects of Gefitinib on the cell proliferation and the expression of epidermal growth factor receptor in human lung cancer cell

作　　者：王亚丽[1] 牟晓燕[1] 游力[2] 白小燕[1] 马春燕[2] 

机构地区：[1]山东大学附属山东省立医院呼吸科,山东济南250021 [2]山东大学附属省立医院中心实验室

出　　处：《中国老年学杂志》2010年第6期805-808,共4页Chinese Journal of Gerontology

基　　金：国家留学归国人员基金项目(2003)

摘　　要：目的探讨表皮生长因子受体(EGFR)抑制剂吉非替尼对肺腺癌A549细胞株增殖和EGFR表达的影响。方法不同浓度吉非替尼干预肺癌A549细胞24、48、72h后,倒置相差显微镜观察药物干预后细胞形态学变化;四甲基偶氮唑盐(MTT)比色法测定细胞抑制率;钙依赖性磷脂结合蛋白(AnnexinⅤ)/碘化丙啶(PI)法和Hoechst33258染色法检测细胞凋亡;流式细胞仪检测药物作用周期;免疫荧光检测EGFR蛋白表达情况;实时荧光定量PCR检测EGFRmRNA的表达情况。结果各干预组细胞出现空泡和颗粒,细胞碎片增多,且随吉非替尼剂量增加和干预时间的延长日益明显。吉非替尼明显抑制了A549细胞的生长,呈时间、剂量依赖性。吉非替尼组细胞凋亡率明显高于正常对照组(P<0.01),S期细胞比例明显减少(P<0.01),G0/G1期细胞比例明显增加(P<0.01),EGFRmRNA和蛋白表达明显减弱(P<0.01)。结论吉非替尼显著抑制A549细胞生长,可能机制是促进凋亡、增强G0～G1期阻滞和进一步下调活化的EGFRmRNA和蛋白的表达。Objective To explore the effects of Gefitinib on proliferation and the expression of epidermal growth factor receptor （EGFR） in lung cancer A549 cell.Methods A549 cells were treated with different concentrations of Gefitinib for 24,48,72 hours.Cellular morphological changes were observed under an inverted microscope.Cell inhibition rate was detected by methyl thiazolyl tetrazolium （MTT）.The apoptosis rate was measured by Annexin V/PI and Hoechst33258 staining method.The cell cycle was detected by the flow cytometer,and the expression of EGFR was determined by immunofluorescence and real-time RT-PCR.Results A great quantity of granules and vacuolus were observed in the intervention groups,also cell debris was increased in a dose-and time-dependent manner.Gefitinib induced a dose-and time-independent growth inhibition by MTT.Higher rate of apoptosis（P〈0.01）,G0/G1 stage cell ratio（P〈0.01,the lower rate of S stage cell proportion（P〈0.01）and the lower EGFR expression （P〈0.01） were present in treatment group of Gefitinib,compared with normal control group.Conclusions Gefitinib can significantly inhibit the growth of A549 cell,possibly by promoting apoptosis,G0-G1 arrest,and down-regulation of EGFR expression.

关 键 词：吉非替尼 凋亡 荧光定量RT-PCR 表皮生长因子受体 

分 类 号：R734[医药卫生—肿瘤]