南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (08): 1110-1113.

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孕激素对人子宫内膜癌细胞系Ishikawa影响的蛋白质组学研究

张瑜； 张怡； 林秋华；   

  1. 中南大学湘雅医院妇产科； 中南大学湘雅二医院妇产科 湖南长沙410008；
  • 出版日期:2006-08-20 发布日期:2006-08-20
  • 基金资助:
    湖南省卫生厅课题(B2004-022)~~

Progesterone-modulated proteins in human endometrial cancer cell line Ishikawa

ZHANG Yu1, ZHANG Yi1, LIN Qiu-hua2 1Department of Obstetrics and Gynecology, Xiangya Hospital, 2Department of Obstetrics and Gynecology, Second Xiangya Hospital, Central South University, Changsha 410078, China   

  1. 中南大学湘雅医院妇产科； 中南大学湘雅二医院妇产科 湖南长沙410008；
  • Online:2006-08-20 Published:2006-08-20

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摘要： 目的寻找并鉴定人子宫内膜癌细胞系Ishikawa在孕激素处理后表达发生变化的蛋白质,为阐明孕激素治疗子宫内膜癌的作用机制提供线索。方法利用固相梯度双向凝胶电泳技术建立药物处理前后细胞总蛋白双向凝胶电泳图谱,经PDQuest软件分析,选取差异表达的蛋白质点进行基质辅助激光解吸电离飞行时间质谱分析,获得肽质量指纹图谱,Mascot软件搜索NCBInr数据库鉴定蛋白质。结果经双向凝胶电泳和图像分析,筛选出41个差异表达蛋白点,其中23个在药物处理后表达上调,18个表达下调。经质谱鉴定的蛋白包括7个孕激素处理后表达上调的蛋白:RAB6A蛋白、splicingfactor3a、儿茶酚氧位甲基转移酶、谷胱甘肽硫转移酶、角蛋白8、CCT6A、AHA1;1个在孕激素处理后表达下调:肽基脯氨酸异构酶A。结论建立了孕激素处理前后的Ishikawa细胞双向蛋白质电泳图谱,初步鉴定了部分与孕激素治疗相关的蛋白,发现孕激素通过干预雌激素的代谢、调节细胞骨架蛋白和细胞内泡膜转运相关蛋白以及分子伴侣蛋白表达等多条途径发挥作用,为进一步了解孕激素作用机制,改善孕激素的疗效提供了新线索。 

Abstract: Objective To identify proteins associated with growth inhibitory effects of progesterone in Ishikawa endometrial adenocarcinoma cell line (Ishikawa). Methods The total cellular proteins of the control and megestrol acetate (MA)-treated Ishikawa cells were separated by two-dimensional gel electrophoresis. After colloidal Coomassie G-250 staining and scanning, the gel images were analyzed by PDQuest software. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and Mascot software were used to identify the differentially expressed protein. Results and Conclusions Well-resolved and reproducible 2-dimensional patterns of both control and MA-treated Ishikawa cells were obtained. Forty-one proteins displayed differential expression after MA treatment, among which 23 were up-regulated and 18 down-regulated. Peptide mass fingerprints (PMF) by MALDI-TOF-MS analysis and search in NCBInr resulted in the identification of 8 proteins. The up-regulated proteins identified were catechol-O-methyltransferase (COMT), glutathione S-transferase, keratin 8, splicing factor 3a (subunit 3), chaperonin containing TCP1 (subunit 6A isoform a, CCT6A), RAB6A protein and AHA1. The down-regulated protein was peptidyl prolyl isomerase A (isoform 1). These findings can be helpful in further investigation of the molecular mechanism of progesterone. 

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