人绒癌耐药细胞株JAR/MTX蛋白质组双向凝胶电泳图谱的建立  

Establishment of two-dimensional gel electrophoresis profiles of proteome from drug-resistant choriocarcinoma cell line JAR/MTX

作　　者：马海鸥[1] 李巨[2] 苏畅[3] 

机构地区：[1]吉林大学第二医院妇产科 [2]沈阳解放军202医院妇产科 [3]吉林大学第一医院妇产科,吉林长春130021

出　　处：《中国妇幼保健》2009年第9期1268-1270,共3页Maternal and Child Health Care of China

摘　　要：目的:建立人绒癌耐药细胞株JAR/MTX蛋白质组双向凝胶电泳图谱。方法:用含10%胎牛血清的RPMI-1640培养液培养JAR/MTX细胞,胰酶消化后收集细胞,加入细胞裂解液使细胞充分裂解,离心后取上清液进行双向凝胶电泳和考马斯亮蓝染色,应用PDQuest图像分析系统进行凝胶图谱分析,从凝胶图中选取1个分离较好的蛋白质点,应用四极杆-飞行时间质谱(Q-TOF-MS)和数据库搜索鉴定蛋白质。结果:获得了背景清晰、分辨率和重复性较好的双向凝胶电泳图谱;在17cmpH3～10IPG胶条上可分离到(810±45)个重复性及分辨率均较好的蛋白位点,蛋白斑点的匹配率为80.2%;1个蛋白点通过质谱分析和数据库检索得到了初步的鉴定。结论:建立了人绒癌耐药细胞株JAR/MTX双向凝胶电泳参考图谱,为其蛋白质组学的进一步研究奠定了基础。Objective： To establish two - dimensional gel eleetrophoresis profiles of proteome from drug - resistant chorioeareinoma cell line JAR/MTX. Methods： JAR/MTX cells were cultured in RPM11640 culture medium containing 10% tetal bovine serum, then har- vested after digestion by trypsin, and lysed in lysis buffer. After centrifugation, supernatant was separated by isoelectrie focusing （IEF） , equilibrated and run on SDS polyacrylamide gel electrophoresis （SDS -PAGE） , then stained by Coomassie brilliant blue.. A well -seperated protein spot was selected from the gel by PDQuest analysis system. Hybrid Quadrupole Time of Flight Mass Spctrometer （ Q - TOF - MS）, peptide mass fingerprinting （PMF） and database searching were used to identify the protein spot. Results： Clear, well -resolved, reproducible two - dimensional gel electrophoresis profiles of proteome in JAR./MTX cells were obtained. （810 ＋ 45 ） protein spots were detected in the two - dimensional gel electrophoresis profiles, of proteome on 17 cm IPG strip （ pH 3 ~ 10） of JAR/MTX cell proteome separated. A match rate of 80. 2% of the spots between gels was obtained. One of these proteins was identified by mass spectrometry and database queries. Conclusion： The two - dimensional gel electrophoresis profiles of proteome from JAR/MTX ceils are established and lay the foundation of further proteomics research of JAR/MTX cells.

关 键 词：JAR/MTX细胞 双向凝胶电泳 蛋白质组 

分 类 号：R71[医药卫生—妇产科学] Q51[医药卫生—临床医学]