表皮生长因子受体抑制剂吉非替尼诱导人表皮角质细胞凋亡的研究  

EGFR inhibitor gefitinib induced keratinocyte apoptosis

作　　者：赵成利[1] 蓝飞晓[1] 谭红梅[1] 匡斌[1] 邓国三[1] 许鹏[2] 王文 郭爱林[2] 

机构地区：[1]广东省医学科学院广东省人民医院整形外科 [2]广东省医学科学院广东省人民医院医学研究中心 [3]广州澳轵朗生物技术有限公司

出　　处：《实用医学杂志》2012年第1期33-36,共4页The Journal of Practical Medicine

基　　金：广东省科技计划项目(编号:93047)

摘　　要：目的:探讨表皮因子受体抑制剂吉非替尼对分离培养的人表皮角质细胞的生长抑制及其诱导凋亡的作用机制,为探寻其对皮肤的毒副作用机制奠定基础。方法:采用分散酶分离表皮细胞,无血清培养法进行表皮细胞的培养,MTT法检测吉非替尼对角质细胞生长的影响,Annexin V-FITC/7-AAD双染流式细胞仪检测细胞凋亡,Western blotting检测Bcl-2、bim的变化,采用caspase-3试剂盒检测caspase-3活性的变化。结果:以分散酶分离法成功得到8例人角质细胞,吉非替尼对人角质细胞生长有显著的抑制作用,并能显著诱导细胞发生凋亡,其作用呈明显的量效依赖性。吉非替尼能明显提高bim表达水平,促进caspase-3的活性。结论:吉非替尼通过调节Bcl-2家族蛋白及激活caspase-3诱导人角质细胞凋亡,抑制角质细胞增殖,这可能是其皮肤毒性的分子生物学基础。Objective To investigate the effect of EGFR inhibitor--gefitinib on inhibiting proliferation and inducing apoptosis of human keratinocytes in vitro and explore the underlying mechanism involved in gefitinib-induced skin toxicity. Methods Keratinocytes were separated by dispase and cultured in serum-free culture medium. The MTT test was used to detect the effect of gefitinib on keratinocytes growth and flow cytometrie analysis with Annexin V- FITC/7-AAD was used to detect the apoptosis of cells. And then Western blotting was performed to analyze the expression of bim and Bcl-2. Caspase-3 activity was detected with Caspase-3 ELISA kits. Results Gefitinib showed obviously suppressed effect on keratinocytes growth and induced apoptosis of keratinocytes in a dose-dependent manner.Gefitinib increased expression level of pro-apoptotie protein-bim obviously, followed by activiting of Caspase-3. Conclusion The underlying mechanism of gefitinib-induced skin toxicity may be that gefitinib induced keratinocytes apoptosis and suppressed keratinocytes growth via the regulation of proteins in Bcl-2 family and Caspase-3 activity.

关 键 词：受体 表皮生长因子 角皮细胞 凋亡 吉非替尼 

分 类 号：R965[医药卫生—药理学]