塞来昔布抑制人肺腺癌细胞的增殖和表皮生长因子受体的表达  

The Effects of Celecoxib on the Cell Proliferation and Expression of Epidermal Growth Factor Receptor in Human Lung Cancer A549 Cells

作　　者：王亚丽[1] 牟晓燕[2] 魏启宏[1] 吴峰[1] 白小燕[2] 刘庆亮[2] 

机构地区：[1]扬州市第一人民医院呼吸科,江苏省扬州市225002 [2]山东大学附属省立医院

出　　处：《中国肿瘤临床》2011年第12期708-711,共4页Chinese Journal of Clinical Oncology

基　　金：山东省自然科学基金(编号:ZR2009CM1250)资助~~

摘　　要：目的:探讨不同浓度的环氧化酶-2抑制剂塞来昔布(Celecoxib)在不同时间点对肺腺癌A549细胞株增殖和表皮生长因子受体(EGFR)表达的影响。方法:人肺腺癌A549细胞株培养于含10%胎牛血清RPM11640培养基中。实验细胞分组如下:A组,正常对照;B组,Celecoxib(12.5 μmol/L);C组Celecoxib(25 μmol/L);D组Celecoxib(50 μmol/L),E组Celecoxib(75 μmol/L),均以RPMI1640培养液配置。使用不同浓度塞来昔布(12.5、25、50和75 μmol/L)处理肺癌A549细胞24、48、72 h后,四甲基偶氮唑盐比色法(MTT)测定细胞增殖抑制率;Annexin Ⅴ/PI染色法与Hoechst33258染色法检测细胞凋亡率;流式细胞仪检测药物作用周期;Real-time RT-PCR法检测EGFR mRNA的表达情况。结果:塞来昔布明显抑制了A549细胞的生长,呈时间、剂量依赖性。塞来昔布组细胞凋亡率明显高于正常对照组(P<0.01),且呈剂量依赖性,S期细胞比例明显减少(P<0.01),G_0/G_1期细胞比例明显增加(P<0.01),提示塞来昔布能将大多数A549细胞阻滞于G_0/G_1期。塞来昔布组细胞EGFR mRNA表达明显减弱(P<0.05),且呈浓度依赖性,各浓度间差异具有统计学意义(P<0.05)。结论:基来昔布显著抑制A549细胞生长,可能机制是通过促进凋亡、增强G_0/G_1期阻滞、下调细胞中EGFR mRNA的表达,从而为应用基来昔布治疗肺腺癌,以及塞来昔布和表皮生长因子受体抑制剂联用提供了一定的实验依据。Objective： To explore the effects of celecoxib on the proliferation and expression of epidermal growth factor receptor （ EGFR ） inhibitor in lung cancer A549 cells at different concentrations and different time points. Methods： A549 cells were cultured in RPMI-1640 and divided into five groups： normal control group; 12.5μmol/L celecoxib group; 25μmol/L celecoxib group; 50 μmol/L celecoxib group; and 75 μmol/L celecoxib group. The cells were treated with celecoxib in different doses （ 12.5, 25, 50, and 75 μmol/L ） and for different periods （ 24, 48, and 72 hours ）. Cell inhibition rate was detected by methyl thiazolyl tetrazolium （ MTT ）. The apoptosis rate was measured using the Annexin V/PI and Hoechst 33258 staining method. The cell cycle was detected by flow cytometry, and the expression of EGFR mRNA was determined through real-time reverse transcriptase polymerase chain reaction. Results： Celecoxib induced a dose- and time-dependent growth inhibition in the intervention groups, as shown by MTT assay. Higher apoptosis rates （ P 〈 0.01 ） and G0/G1 stage cell ratios （ P〈 0.01 ）, and lower rates orS stage cell proportion （ P〈 0.01 ）, and EGFR mRNA expression （ P〈 0.01 ） were observed in the celecoxib treatment groups compared with the normal control group. Conclusion： Celecoxib significantly inhibits the growth ofA549 cell, possibly by promoting apoptosis, G0-G1 arrest, and downregulation of EGFR mRNA expression. Celecoxib has potential application in lung cancer treatment. Our study provides a new therapeutic use for celecoxib combined with EGFR inhibitors.

关 键 词：塞来昔布 凋亡 表皮生长因子受体 环氧化酶-2 

分 类 号：R734.2[医药卫生—肿瘤]