流式细胞仪分析癌细胞对肥大细胞募集的影响

The Analysis of the Influence of Flow Cytometry for Cancer Cells on the Recruitment of Mast Cells

目的:研究食管癌细胞迁移到小鼠腹腔时肿瘤细胞周边微环境的变化。方法:将食管癌细胞株EC109和/或诱导试剂植入小鼠腹腔,利用组织化学的方法、荧光标记的肥大细胞蛋白酶和流式细胞术,我们在小鼠模型观察肠组织的形态和腹腔的MC亚型的变化。结果:胰酶导致小鼠肠道平滑肌层和黏膜下层的组织增厚,细胞间隙的增加可能有益于MC在组织移动或迁移进入腹腔;它引起小鼠腹腔液的总MC增加,MCC亚型的相对比例增加,MCT亚型减少。EC109细胞不能明显地改变小鼠肠道组织的形态,但它显著地引起腹腔MCT亚型的相对比例增加。结论:根据肥大细胞内颗粒的类胰蛋白酶和类糜蛋白酶的差异表达,可证实小鼠的MC亚型;并且不同的诱导物可能影响腹部微环境的变化。目前的研究表明,食道癌细胞可以诱导MCT(含有类胰蛋白酶)亚型迁移到小鼠腹腔,造成肿瘤细胞周围的内部环境的变化。

Objective： To study the change of neighboring microenvironment in cancer cells during the esophageal carcinoma cells migrating into mouse abdominal cavity. Methods： After implanting esophageal carcinoma cells line EC109 and / or inducing reagent into mouse abdominal cavity,we can observe the morphology of intestine organization and the change of MC subtypes of abdominal cavity in mouse models by using histochemical method,fluorescence labeling of mast cells protease and flow cytometry. Results： The smooth muscle layer and submucosa of mouse intestinal tissue became thicken because of tyrisin,and the increase of intercellular gap could be beneficial for MC to migrate to tissue or into abdominal cavity. It aroused the increase of total MC in mouse abdominal fluid and the relative percentage of MCCsubtype,while the MCTsubtype decreased. Although EC109 cells could not obviously change the morphology of mouse intestinal tissue,it dramaticlly results in the increase of relative percentage of MCTsubtype in abdominal cavity. Conclusions： According to the differences between the expression of tryptase and chymase in mast cell granules,the mouse MC subtypes can be confirmed,in addition,abdominal microenvironment changes could aslo be affected by various inducer. The present study suggests that esophageal cancer cells can make MCT（ containning tryptase） subtype migrate into mouse abdominal cavity,which cause the change of the internal environment surrounding the cancer cells.