重组人信号素3A抑制胃癌血管生成及其机制  

Inhibitory effect of recombinant human semaphorin 3A on angiogenesis of gastric cancer and the associated mechanisms

作　　者：冯品 范文静 刘镭[2] 许倩[3] 李玉红 左彦珍[5] 周搏 赵向阳[7] FENG Pin;FAN Wen-jing;LIU Lei;XU Qian;LI Yu-hong;ZUO Yan-zhen;ZHOU Bo;ZHAO Xiang-yang(Department of Pathology,Chengde Medical University,Hebei Chengde 067000,China;Department of Immunology,Chengde Medical University,Hebei Chengde 067000,China;Basic Medical Institute,Chengde Medical University,Hebei Chengde 067000,China;Academic Affairs Office,Chengde Medical University,Hebei Chengde 067000,China;Department of Pharmacology,Chengde Medical University,Hebei Chengde 067000,China;Department of Neurology,981 Hospital of Chinese PLA,Hebei Chengde 067000,China;Department of General Surgery,981 Hospital of Chinese PLA,Hebei Chengde 067000,China)

机构地区：[1]承德医学院病理学教研室,河北承德067000 [2]承德医学院免疫学教研室,河北承德067000 [3]承德医学院基础医学研究所,河北承德067000 [4]承德医学院教务处,河北承德067000 [5]承德医学院药理学教研室,河北承德067000 [6]中国人民解放军第981医院神经内科,河北承德067000 [7]中国人民解放军第981医院普通外科,河北承德067000

出　　处：《解剖学报》2020年第2期220-227,共8页Acta Anatomica Sinica

基　　金：河北省高校重点学科建设项目(冀教高〔2013〕4号);河北省高校百名优秀创新人才支持计划Ⅲ项目(SLR2017018)。

摘　　要：目的探讨信号素3A(Sema3A)及其受体神经激肽-1(NRP-1)在胃癌中的表达及其与微血管密度(MVD)的相关性;并探讨重组人Sema3A对胃癌血管生成的影响及其相关的机制。方法选取40例手术切除胃癌组织及其癌旁正常组织,免疫组织化学法检测组织中Sema3A、NRP-1的表达及MVD。ELISA检测胃癌患者组及正常对照组血清Sema3A的表达水平。Western blotting检测5株胃癌细胞系(MGC-803、HGC-27、MKN-28、SGC-7901、MKN-45)、人正常胃黏膜细胞(GES-1)中Sema3A和NRP-1的表达。应用Transwell小室构建非接触式体外共培养体系,小管形成实验初步研究不同浓度重组人Sema3A对胃癌血管生成的影响;Western blotting检测共培养体系中血管内皮生长因子受体2(VEGFR2)、NRP-1的表达水平。结果胃癌组织、细胞和患者血清中Sema3A的表达显著低于对照组(P<0.05),胃癌组织和MKN-28细胞中的NRP-1表达明显升高(P<0.05),两者均与胃癌TNM分期相关(P<0.05),且Sema3A与微血管密度存在负相关(P<0.05)。在体外共培养体系中,重组人Sema3A处理组人脐静脉内皮细胞(HUVEC)小管形成能力下降,且具有浓度依赖性;重组人Sema3A可下调VEGFR2蛋白的表达。结论Sema3A在胃癌组织、细胞和患者血清中均表达降低,与微血管密度成负相关,重组人Sema3A具有体外抑制胃癌血管生成的作用,可能与下调VEGFR2蛋白表达有关。Objective To investigate the expression of semaphorin 3A(Sema3A)and its receptor neuropilin-1(NRP-1)in gastric cancer and its correlation with microvessel density(MVD),and then to explore the effect of recombinant human Sema3A on angiogenesis of gastric cancer and the associated mechanisms.Methods Forty cases of gastric cancer tissues and its corresponding adjacent normal tissues were used to detecte the expression of Sema3A,NRP-1 and MVD in tissues by immunohistochemistry method.The expression level of Sema3A in serum of gastric cancer patient group and normal control group were measured by Enzyme-linked immuno-sorbent assay(ELISA).Western blotting was used to detect the expression of Sema3A and NRP-1 in five gastric cancer cell lines(MGC-803,HGC-27,MKN-28,SGC-7901,MKN-45)and human gastric mucosal epithelial cell(GES-1).Transwell chamber was used to construct non-contact in vitro co-culture system,in which the effects of different concentrations of recombinant human Sema3A on angiogenesis in gastric cancer were analyzed by tube formation assay preliminarily.The expression levels of vascular endothelial growth factor receptor 2(VEGFR2)and NRP-1 in co-culture system were detected by Western blotting.Results The expression levels of Sema3A in gastric cancer tissues,cell lines and patient serum were significantly lower than that in the control group(P<0.05),while the expression of NRP-1 in gastric cancer tissues and MKN-28 cells was significantly increased,and both of them were associated with TNM staging of gastric cancer(P<0.05).In vitro co-culture system,The tube forming abilities of human umbilical vein endothelial cell(HUVEC)were decreased in recombinant human Sema3A treated group,and this phenomenon was concentration dependent.The expression of VEGFR2 protein was down-regulated by recombinant human Sema3A.Conclusion The expression of Sema3A was decreased in gastric cancer tissues,cell lines and patient serum,and negatively correlated with microvessel density.The recombinant human Sema3A could inhibit the angio

关 键 词：胃癌 信号素3A 脐静脉内皮细胞 共培养体系 小管形成实验 免疫印迹法 人 

分 类 号：R656.6[医药卫生—外科学]