作　　者： ; ; ; ; ; ; ;

机构地区： 汕头大学医学院

出　　处： 《中国实验动物学报》 2012年第5期62-68,I0005,共8页

摘　　要： 目的建立小鼠肥大细胞(mast cell,MC)迁移模型,探讨化疗药物三氧化二砷对人食管癌EC109细胞株生长的杀伤机理。方法利用肥大细胞的特征性蛋白酶抗体及其免疫荧光标记MC和PI标记EC109细胞内DNA;以流式细胞术分析小鼠腹腔液中肥大细胞各亚型的百分率及肿瘤细胞周期变化;使用激光扫描共聚焦显微镜显示肥大细胞内分泌颗粒的分布。并通过组织化学方法,观察各种诱导处理后小鼠肠组织肥大细胞由肠道向腹腔移动的变化。结果①根据流式细胞仪点图分布分析,将小鼠肥大细胞分为:类胰蛋白酶阳性、类糜蛋白酶阴性(MC T);类糜蛋白酶阳性、类胰蛋白酶阴性(MC C)和类胰蛋白酶阳性、类糜蛋白酶阳性(MC TC)三种亚型,且T型MC明显多余TC型和C型(P<0.05);以共聚焦显微镜显示三种亚型的MC均含有丰富的分泌颗粒并分布于细胞膜内侧,为其出芽突起形成储备的状态。②经组织切片观察到诱导处理后小鼠肠组织MC由肠道向腹腔移动,且胰酶对MC的诱导作用大于食管癌细胞和As2O3。③经诱导迁移入腹腔的MC可能与癌细胞周期由S期向G2/M期跨越相关;As2O3能延迟食管癌细胞的G0/G1期,阻碍细胞向S期跨越,从而抑制食管癌细胞的生长。结论食管癌细胞移入小鼠腹腔,主要诱导T型MC参与免疫反应。在生物机体内环境(这里指MC影响)的条件下,As2O3对肿瘤细胞生长的作用主要表现为促使癌细胞周期的G0/G1期向S期跨越延迟,或G2/M期进入细胞分裂的延迟。 Objective To establish an experimental mouse model of mast cell（MC） migration,and explore the killing mechanism of chemotherapeutic drug As2 O3 on human esophageal carcinoma EC109 cell line.Methods Mouse mast cell（MC） subsets were defined on the basis of neutral protease composition and immunofluorescence staining,and the distribution image of DNA content in esophageal carcinoma cells was analyzed by propidium iodide labeling and flow cytometry.Secretory granules in MC were observed by laser scanning confocal microscopy.The MC migration from the gut to peritoneal cavity after induction treatments were observed by immunohistochemistry.Results ① According to the distribution of flow cytometric dot plot analysis,the mouse MCs were divided into three immunophenotypes： tryptase-positive and chymase-negative MCs（MC T）;chymase-positive and tryptase-negative MCs（MC C）,and both tryptase-positive and chymase-positive MCs（MC TC）.The amount of MC T cells is apparently more than that of MC TC and MC C cells（P0.05）.The laser scanning confocal microscopic observation revealed that all the three subtypes of MCs contained profuse secretory granules distributed in the inner membrane just ready for budding into the extracellular space.② The mouse MCs migrated from the intestinal tissue into the peritoneal cavity after the induction treatment.The trypsin-induced MC migration was more intensive than that induced by esophageal cancer cells and As2 O3.③ The MC migration into peritoneal cavity may be related to cancer cell cycle transition from S phase to G2/M phase.As2 O3 could delay esophageal cancer cell cycle in G0/G1 phase and impeded cells cycle into S phase,thus inhibiting the growth of esophageal cancer cells.Conclusions The esophageal cancer cells injected into the peritoneal cavity of mice mainly induce T MCs to be involved into immune responses.At the presence of MCs in vivo,the effect of As2 O3 on tumor cell growth is mainly to delay the cell cycle from G0/G1 phase to S phase,or delay the tran

关 键 词： 肥大细胞 类胰蛋白酶 食管癌 细胞周期 三氧化二砷 小鼠

领　　域： [生物学]