中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
10期
699-702
,共4页
夏汝山%顾静%陶诗沁%杨莉佳
夏汝山%顧靜%陶詩沁%楊莉佳
하여산%고정%도시심%양리가
毛乳头细胞%分泌蛋白质类%电泳,凝胶,双向%MALDI-TOF质谱
毛乳頭細胞%分泌蛋白質類%電泳,凝膠,雙嚮%MALDI-TOF質譜
모유두세포%분비단백질류%전영,응효,쌍향%MALDI-TOF질보
Dermal papilla cells%Secretory proteins%Electrophoresis,gel,two-dimensional%MALDI-TOF mass spectrometry
目的 了解毛乳头细胞在凝集生长状态下分泌性蛋白组的表达.方法 以凝集和非凝集生长的毛乳头细胞为研究对象,分别制备分泌性蛋白质,以双向凝胶电泳技术和PDQuest软件分析二者之间的蛋白图谱的差异,通过基质辅助激光解吸电离飞行时间串联质谱(MLDI-TOF-MS)鉴定表达差异的蛋白质,通过蛋白质数据库NCBInr检索分析.结果 建立了重复性好、分辨率高的双向电泳图谱;在凝集生长和非凝集生长的毛乳头细胞分泌性蛋白质中分别检测到(1 134±52)个和(1 078±36)个蛋白点,大多匹配.按照差异量在5倍以上的标准,二者存在差异蛋白质28个,经过MLDI-TOF-MS质谱鉴定出10种差异表达蛋白点,其中8种蛋白质表达上调,分别为RhoGDP分离抑制剂1、细丝蛋白A、胱抑素C、纤维连接蛋白、亲环素A、前胶原C端蛋白酶增强子、组织金属蛋白酶抑制剂、组织金属蛋白酶-2抑制剂.2种蛋白质表达下调,为神经肽h3和基质金属蛋白酶-3金属蛋白酶组织抑制剂-1复合物复合物.结论 凝集生长和非凝集生长毛乳头细胞差异蛋白主要涉及信号通路、细胞增殖与分化、细胞外基质合成及降解等功能.
目的 瞭解毛乳頭細胞在凝集生長狀態下分泌性蛋白組的錶達.方法 以凝集和非凝集生長的毛乳頭細胞為研究對象,分彆製備分泌性蛋白質,以雙嚮凝膠電泳技術和PDQuest軟件分析二者之間的蛋白圖譜的差異,通過基質輔助激光解吸電離飛行時間串聯質譜(MLDI-TOF-MS)鑒定錶達差異的蛋白質,通過蛋白質數據庫NCBInr檢索分析.結果 建立瞭重複性好、分辨率高的雙嚮電泳圖譜;在凝集生長和非凝集生長的毛乳頭細胞分泌性蛋白質中分彆檢測到(1 134±52)箇和(1 078±36)箇蛋白點,大多匹配.按照差異量在5倍以上的標準,二者存在差異蛋白質28箇,經過MLDI-TOF-MS質譜鑒定齣10種差異錶達蛋白點,其中8種蛋白質錶達上調,分彆為RhoGDP分離抑製劑1、細絲蛋白A、胱抑素C、纖維連接蛋白、親環素A、前膠原C耑蛋白酶增彊子、組織金屬蛋白酶抑製劑、組織金屬蛋白酶-2抑製劑.2種蛋白質錶達下調,為神經肽h3和基質金屬蛋白酶-3金屬蛋白酶組織抑製劑-1複閤物複閤物.結論 凝集生長和非凝集生長毛乳頭細胞差異蛋白主要涉及信號通路、細胞增殖與分化、細胞外基質閤成及降解等功能.
목적 료해모유두세포재응집생장상태하분비성단백조적표체.방법 이응집화비응집생장적모유두세포위연구대상,분별제비분비성단백질,이쌍향응효전영기술화PDQuest연건분석이자지간적단백도보적차이,통과기질보조격광해흡전리비행시간천련질보(MLDI-TOF-MS)감정표체차이적단백질,통과단백질수거고NCBInr검색분석.결과 건립료중복성호、분변솔고적쌍향전영도보;재응집생장화비응집생장적모유두세포분비성단백질중분별검측도(1 134±52)개화(1 078±36)개단백점,대다필배.안조차이량재5배이상적표준,이자존재차이단백질28개,경과MLDI-TOF-MS질보감정출10충차이표체단백점,기중8충단백질표체상조,분별위RhoGDP분리억제제1、세사단백A、광억소C、섬유련접단백、친배소A、전효원C단단백매증강자、조직금속단백매억제제、조직금속단백매-2억제제.2충단백질표체하조,위신경태h3화기질금속단백매-3금속단백매조직억제제-1복합물복합물.결론 응집생장화비응집생장모유두세포차이단백주요섭급신호통로、세포증식여분화、세포외기질합성급강해등공능.
Objective To study the expression ot secreted proteins in aggregated dermal papilla cells (DPCs).Methods DPCs were isolated from human scalp tissue and subjected to primary culture and subculture.Aggregated and non-aggregated DPCs served as the subject of this study.Secreted proteins were prepared from these cells and subjected to two-dimensional polyacrylamide gel electrophoresis.Differentially expressed proteins were screened by the PDQuest image analysis software.Protein spots were digested and identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry,and finally analyzed using the National Center for Biotechnology Information (NCBI) non-redundant (Nr) protein database.Results Two-dimensional electrophoresis maps with good repeatability and high resolution were established.Image analysis of 2-D gels revealed that the average number of detected protein spots was 1 134 ± 52 and 1 078 ± 36 in aggregated and nonaggregated DPCs respectively,and the majority of these protein spots were matched between aggregated and nonaggregated DPCs.Twenty-eight protein spots showed more than 5-fold difference between the two groups of cells,and 10 proteins were preliminarily identified as differentially expressed proteins by peptide-mass fingerprinting.Of these differentially expressed proteins,8 proteins including Rhogdi 1,filamin A,cystatin C,fibronectin,cyclophilin A,procollagen C proteinase enhancer 1,tissue inhibitor of metalloproteinase and tissue inhibitor of metalloproteinase-2 were up-regulated,and 2 proteins including neuropolypeptide h3 and matrix metalloproteinase-3/tissue inhibitor of metalloproteinase-1 complex were down-regulated in aggregated DPCs compared with non-aggregated DPCs.Conclusions Differentially expressed proteins between aggregated and non-aggregated DPCs are mainly implicated in cell signaling pathway,cellular proliferation and differentiation,extracellular matrix synthesis and degradation,and so on.
