hepacam蛋白通过exosomes途径分泌到细胞外 - 第三军医大学学报.doc

HepaCAM蛋白通过exosomes途径分泌到细胞外并抑制肿瘤细胞增殖 刘安全1 吴小候1罗春丽2 (400016，重庆，重庆医科大学第一医院泌尿外科1，重庆医科大学附属第一医院泌尿外科，.重庆医科大学医学检验系2) 【摘要】：目的 探讨肝细胞黏附分子（hepatocyte cell adhesion molecule,hepaCAM）在肾癌786-0细胞中。将携带hepaCAM基因的重组质粒瞬时转染786-0细胞,鉴定hepaCAM基因在786-0中的表达;。结果上述结,hepaCAM蛋白可能通过 Is Secreted by Human Tumor Cells through exosomes pathway and Inhibit Proliferation of Tumor Cells Liu Anquan , WU Xiaohou ,Luo Chunli, （Chongqing 400016，China，Department of Urology，First Hospital，Chongqing Medical University1，Department of Clinical Laboratory Medicine） Abstract] Objective This study was to investigate that whether hepatocyte cell adhesion molecule(hepaCAM)is actively secreded from renal cell cancer by a pathway involving exosomes and wether hepaCAM could inhibit proliferation of renal cell cancer. Methods a recombinant adenovirus vectors plasmid containing hepaCAM was transfected into 786-0 cells.The expression of hepaCAM was detected by western blot.The transfected cells was treated with monensin and 5-(N-N-Dimetyl)-amiloride hydrochloride (DMA),and then extracellular culture medium and exosomes were used to investigate the present of hepaCAM. Results HepaCAM adenovirus vectors plasmid was successfully constructed and stably transfected into 786-0 cells. Monensin enhance release of hepaCAM and DMA inhibit release of hepaCAM from transfected 786-0 cell. Exosomes puritied from transfected 786-0 cells inhibit the proliferation of 786-0 cells.Conclusion HepaCAM is secreted from renal cell cancer by a pathway involving exosomes. HepaCAM is presten in exosomes collected form 786-0 cells transfected with hepaCAM adenovirus plasmid. Exosomes collected from 786-0 cells transfected with hepaCAM adenovirus plasmid inhibit proliferation of renal cell cancer. [Key words] HepaCAM;exosomes;secreted，proliferation HepaCAM是由新加坡沈莎莉教授于2004年发现的一种是新基因，它编码的是一个免疫球蛋白样跨膜糖蛋白，是粘附分子中免疫球蛋白超家族，该基因在肿瘤细胞内成低表达3，过表达后可明显抑制肿瘤细胞增殖4。肿瘤细胞内存在大量肿瘤相关蛋白，对肿瘤细胞的发生发展起着重要的作用，同时肿瘤细胞可以通过多种方式将多种活性因子及蛋白分泌到细胞外并发挥一定的作用，如HSP60 5.6、HSP70 7.8等，HS