癌源外泌体的制备及其对胃癌细胞STK15表达的影响  

Identification and isolation of exosomes derived from tumor and their effects on STK15 expression on gastric cancer cells

作　　者：杨勇 陈霖 杨宏新 李磊 于建 何源哈达 杨晨 徐伟 高艳伟 高维实 YANG Yong;CHEN Lin;YANG Hongxin;LI Lei;YU Jian;HEYUAN Hada;YANG Chen;XU Wei;GAO Yanwei;GAO Weishi(Department of Oncology,Inner Mongolia People's Hospital,Hohhot,Neimenggu 010017,China;School of Basic Medicine,Inner Mongolia Medical University,Hohhot,Neimenggu010059,China;Department of Traditional Chinese Medicine,Yichang People's Hospital,Yichang,Hubei 443000,China;College of Clinical Medicine,Inner Mongolia Medical University,Hohhot 010059,China;Department of Combined Chinese and WesternMedicine,Inner Mongolia People's Hospital,Hohhot,Neimenggu 010017,China)

机构地区：[1]内蒙古自治区人民医院肿瘤外科,呼和浩特010017 [2]内蒙古医科大学基础医学院,呼和浩特010059 [3]湖北省宜昌市第一人民医院中医科,443000 [4]内蒙古医科大学临床医学院,呼和浩特010059 [5]内蒙古自治区人民医院中西医科,呼和浩特010017

出　　处：《重庆医学》2019年第9期1453-1456,1461,共5页Chongqing medicine

基　　金：内蒙古自然科学基金资助项目(2014MS0864);内蒙古肿瘤生物治疗协同创新项目(2016ZLXT008)

摘　　要：目的探讨胃癌来源外泌体对胃癌细胞的增殖作用及其对丝氨酸/苏氨酸激酶15(STK15)表达的影响。方法采用超速梯度离心法从胃癌MGC803细胞培养上清液中提取外泌体,通过透射电镜技术对其超微结构进行鉴定,然后用不同浓度的外泌体(0.05、0.10、0.20、0.50μg/μL)处理胃癌细胞,利用甲基偶氮唑蓝(MTT)法检测其对细胞增殖活性的影响。利用免疫细胞化学方法分析外泌体对胃癌细胞中心体扩增激酶STK15表达的影响。结果胃癌MGC803细胞来源的外泌体具有特征性的盘状结构,由双层膜构成,直径为50～130nm。外泌体具有促进胃癌细胞增殖活性的作用(P<0.05)。随着时间的延长增殖活性也增强,12h细胞增殖活性从0.213上升到0.534(F=44.21,P=0.000),24h细胞增殖活性从0.251上升到0.998(F=66.06,P=0.000),36h细胞增殖活性从0.273上升到1.057(F=435.37,P=0.000),48h细胞增殖活性从0.342上升到1.414(F=195.30,P=0.000),外泌体以剂量依赖性-时间依赖性促进胃癌细胞增殖,其可能通过负载抗原上调STK15表达来影响细胞增殖进程。结论胃癌来源外泌体可能通过调节细胞STK15表达促进胃癌细胞增殖。Objective To discuss the effect of exosomes on proliferation of gastric cancer cells and expression of serine/threonine kinase 15(STK15).Methods Exosomes were extracted from the culture supernatant of MGC803 gastric cancer cells by ultracentrifugal gradient centrifugation,and their ultrastructure was identified by transmission electron microscopy.Gastric cancer cells were then treated with exosomes at different concentrations(0.05,0.10,0.20,0.50μg/μL)The effect of exosomes on the proliferation of gastric cancer cells was examined by MTT assay.Immunohistochemical method was used to test the expression of serine/threonine kinase 15(STK15)in gastric cancer cells.Results The exosomes derived from gastric cancer MGC803 cells had a characteristic plate-like structure consisting of a bilayer membrane with the diameter of about 50-130 nm.Exosoms had the effect of promoting the proliferation acitivity of gastric cancer cells(P<0.05).The cell proliferation activity increased from 0.213 to 0.534 when treated 12 h(F=44.21,P=0.000),from 0.251 to 0.998 when treated 24 h(F=66.06,P=0.000),from 0.273 to 1.057 when treated 36 h(F=435.37,P=0.000),from 0.342 to 1.414 when treated 48 h(F=195.30,P=0.000).Exosomes promoted the proliferation of MGC803 cells in a dose-dependent and time-dependent manner,which might affect the cell proliferation process by regulating the expression of STK15.Conclusion Exosomes can accelerate the proliferation of gastric cancer cells by regulating STK15 expression.

关 键 词：外泌体 胃肿瘤 丝氨酸/苏氨酸激酶15 细胞增殖 

分 类 号：R73-3[医药卫生—肿瘤]