接合物蛋白Crk参与卵巢粘液性上皮癌细胞MCAS的粘液形成.DOC

接合物蛋白Crk在卵巢黏液性上皮癌细胞MCAS黏液形成中的作用 王 辉1，令狐华1，2，徐冬梅1，津田真寿美2，田中伸哉2，長嶋和郎2 （1重庆医科大学附属第一医院妇产科，重庆400016；2日本北海道大学医学研究生院分子细胞病理学教研室，日本 札幌 060 8638） 摘 要： 目的 阐明接合物蛋白Crk表达缺陷对人卵巢黏液性上皮癌细胞株MCAS细胞的黏液产生的影响。 方法 细胞质PAS染色和颗粒排除检测法分别评估细胞内、外黏液产生情况，PAS染色评估裸鼠模型所形成的肿瘤内黏液形成情况。结果 相差显微镜和细胞质PAS染色观察到Crk表达缺陷性MCAS细胞的细胞体增大，细胞质内充满小泡样黏液。颗粒排除检测法发现Crk表达缺陷性MCAS细胞有更多的黏液分泌，表现为细胞外有更多的基质存在，将红细胞颗粒排除在外。裸鼠模型的肿瘤体内也表现出黏液区的增多。 结论 卵巢黏液性上皮癌的癌变过程与黏液产生和分泌模式的改变有关。接合物蛋白Crk可能同时参与了这两个过程。 关键词：卵巢黏液性上皮癌；细胞质PAS染色；颗粒排除检测法；接合物蛋白Crk 中图法分类号: 文献标志码: A The role of adaptor protein Crk in The Mucus Formation in Mucinous Epithelial Ovarian Cancer Cells MCAS WANG Hui1, LINGHU Hua1,2, XU Dong-mei1, TSUDA Masumi2, TANAKA Shinya2; NAGASHIMA Kazuo2 （1Department of Obstetrics and Gynecology, the First Affiliated Hospital ,Chongqing Medical University, Chongqing 400016, China；2Laboratory of Molecular and Cellular Pathology, Hokkaido University Graduate School of Medicine, N15, W7, Sapporo 060 8638, Japan） Abstract Objective The mucus formation was thought to be an indicator for the histological grade of mucinous epithelial ovarian cancer (mEOC). Previously, Crk expression was targeted in the human ovarian mucinous adenocarcinoma cell line MCAS through RNA interference, and Crk knock down cells were established. These cells exhibited decreased tumorigenic potential both in vitro and in vivo. The aim of this study was to investigate if there is any change in the capability of forming mucus in these Crk knock down cells. Methods Cytoplasmic periodic acid Schiff (PAS) staining and particle excluding assay were conducted to assess the mucus formation within and around cells, respectively. Moreover, the amount of mucus formed in tumor lumps from nude mice model was measured following PAS staining. Results The increased mucus production in Crk knockdown mEOC cells (MCAS) was demonstrated by increased number of enlarged cells filled with vacuoles-like mucus observed by phase-contrast microscope and cytoplasmic P