rhPTH(1-34)诱导角质形成细胞蛋白酪氨酸激酶差异表达蛋白的筛选  

Screening for Differentially Expressed Proteins of Protein Tyrosine Kinase in rh PTH(1-34)-induced Keratinocytes

作　　者：卜晓琳[1] 毕新岭[2] 顾军[2] 汪五清 史玉玲[4] 徐晓光[2] 王国江 

机构地区：[1]上海市浦东新区公利医院皮肤科,上海200135 [2]第二军医大学长海医院皮肤科,上海200433 [3]上海市闵行区中心医院皮肤科,上海201100 [4]同济大学附属第十人民医院皮肤科,上海200072 [5]上海市浦东新区周浦医院皮肤科,上海201318

出　　处：《中国皮肤性病学杂志》2016年第3期221-224,238,共5页The Chinese Journal of Dermatovenereology

基　　金：浦东新区卫生系统优秀青年医学人才培养计划(PWRq2013-02);上海市科委生物医药领域产学研合作项目(13DZ1930906)

摘　　要：目的利用蛋白芯片技术,筛选重组人甲状旁腺素(1-34)[rh PTH(1-34)]作用角质形成细胞后,156种蛋白酪氨酸激酶中的差异表达蛋白。方法培养Ha Ca T角质形成细胞株,rh PTH(1-34)诱导细胞,经过总蛋白质提取、BCA蛋白质定量、酪氨酸磷酸化抗体检测、图象和数据采集,筛选出差异蛋白,最后对所得数据进行分析。结果 156种蛋白中,rh PTH(1-34)组与空白对照组比较,20种蛋白表达差异2倍以上,其中16种显著上调,分别为:Csk,Dtk,Eph A4,Eph A5,Eph B2,Eph B3,Erb B4,FAK,Insulin R,LTK,PDGFR-α,ROS,Tie-1,Tie-2,Tyk2,TYRO10;4种显著下调,分别为:EGFR,JAK3,ALK,Eph B1。其中与银屑病相关的EGFR,JAK3,Tyk2,Eph A4主要涉及角质形成细胞增殖、分化,调节T细胞介导的炎症反应。结论 rh PTH(1-34)可影响Ha Ca T细胞内一组蛋白酪氨酸激酶的表达,其中EGFR,JAK3,Tyk2,Eph A4与银屑病的发病机制密切相关,值得进一步深入探讨。Objective To screen the differentially expressed proteins of 156 kinds of protein tyrosine kinase in rh PTH（1-34）-induced keratinocytes using protein array. Methods We cultured keratinocyte cell line Ha Ca T. Cells were divided into two groups,rh PTH（ 1-34） treatment group and blank control group. Total proteins were extracted. BCA protein quality was assessed. Protein tyrosine kinase antibody was detected. The signal images were scanned by a laser scanner and acquired images were analyzed by software. Subsequently,the data of these groups were compared and screened to differentially expressed proteins. Results Among 156 proteins,20 were identified to be differentially expressed between rh PTH（ 1-34）-treated group and control group,including 16 up-regulated proteins（ Csk,Dtk,Eph A4,Eph A5,Eph B2,Eph B3,Erb B4,FAK,Insulin R,LTK,PDGFR-α,ROS,Tie-1,Tie-2,Tyk2,TYRO10） and 4 down-regulated proteins（ EGFR,JAK3,ALK,Eph B1）.EGFR,JAK3,Tyk2,Eph A4,which were associated with psoriasis,were mainly involved in the proliferation and differentiation of keratinocytes and inflammatory reaction of T cells. Conclusion rh PTH（ 1-34） could affect the expression of a set of protein tyrosine kinase in Ha Ca T cells,including EGFR,JAK3,Tyk2 and Eph A4,which were closely related to the pathogenesis of psoriasis.

关 键 词：角质形成细胞 重组人甲状旁腺素(1-34) 蛋白酪氨酸激酶 差异表达蛋白 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]