B7-H3^＋ TEMs在肾透明细胞癌血管生成中的作用  

Experimental study on B7-H3^＋ Tie2 expressing monocytes mediated angiogenesis in dear cell renal cell carcinoma

作　　者：张先云 黄玉华[1] 张光波 姜福金 侯建全[1,2] Zhang Xianyun;Huang Yuhua;Zhang Guangbo;Jiang Fujin;Hou Jianquan(Department of Urology, First Affiliated Hospital of Soochow University, Suzhou 215006, China)

机构地区：[1]苏州大学附属第一医院泌尿外科,215006 [2]江苏省临床免疫研究所 [3]徐州医科大学附属淮安医院泌尿外科

出　　处：《中华泌尿外科杂志》2018年第6期461-466,共6页Chinese Journal of Urology

基　　金：国家自然科学基金（81272839,81472401）

摘　　要：目的探讨B7-H3在Tie2表达单核巨噬细胞（Tie2 expressing monocytes, TEMs）介导肾透明细胞癌（ccRCC）血管生成中的作用。方法2016年4—12月采用流式细胞术检测确诊的20例肾透明细胞癌组织及癌旁正常肾组织中TEMs表面B7-H3表达情况；利用免疫组化染色法检测肾透明细胞癌B7-H3^＋TEMs高表达和B7-H3^＋TEMs低表达者CD34标记的微血管密度之间的差异；将786-O肾癌细胞株与B7-H3^＋TEMs和B7-H3^－TEMs共培养，收集B7-H3^＋TEMs和B7-H3^－TEMs培养上清液作为条件培养基（分别为B7-H3^＋TEMs组和B7-H3^－TEMs组，另外以普通培养基为空白对照组），通过小管形成实验和小鼠主动脉环实验验证B7-H3^＋TEMs对血管形成能力的影响。结果肾透明细胞癌组织中TEMs表面B7-H3分子表达率为（45.10±17.78）%，癌旁正常肾组织的表达率为（10.28±4.28 ）%，差异有统计学意义（P〈0.01）；B7-H3^＋TEMs低表达者微血管密度为76.55±20.80，B7-H3^＋TEMs高表达者微血管密度为103.81±29.28，差异有统计学意义（P=0.027）；小管形成实验结果显示B7-H3^＋TEMs组脐静脉内皮细胞的小管形成数量（55.25±11.48）显著多于B7-H3^－TEMs组（31.34±8.45）和空白对照组（25.00±6.74） （P〈0.001）。小鼠主动脉环实验结果显示B7-H3^＋TEMs组新生血管形成数量（77.35±18.47）显著多于B7-H3^－TEMs组（39.42±8.29）和空白对照组（28.79±7.63） （P〈0.001）。结论B7-H3＋TEMs在肾透明细胞癌的血管生成中可能发挥重要作用，有望成为肾透明细胞癌抗血管生成治疗的有效靶标。ObjectiveTo explore the role of B7-H3 in the Tie2 expressing monocytes （TEMs） mediated angiogenesis of clear cell renal cell carcinoma （ccRCC）.MethodsLevel of B7-H3 expression on TEMs surface was detected by flow cytometry in ccRCC tissues and normal renal tissues, which were obtained from April 2016 to August 2016 from 20 patients. Microvessel density （MVD） labeled by CD34 in high B7-H3^＋ TEMs group and low B7-H3^＋ TEMs group was detected by immunohistochemical examination in ccRCC specimens. B7-H3^＋ TEMs and B7-H3^-TEMs were co-cultured with the 786-O cell lines, and B7-H3＋ TEMs and B7-H3^-TEMs culture supernatants were collected as conditioned medium, then the effect of B7-H3^＋ TEMs on angiogenesis was tested by tubule formation assay and mouse aortic ring assay.ResultsFlow cytometry showed that the frequency of B7-H3 expression on TEMs in ccRCC was （45.10±17.78）%, and the frequency of B7-H3 expression in normal kidney tissues was （10.28±4.28）%. The frequency of B7-H3 expression on TEMs was significantly higher than that in normal renal tissues （P〈0.001）. The MVD of high B7-H3^＋ TEMs group （103.81±29.28） was higher than that of low B7-H3^＋ TEMs group （76.55±20.80） （P=0.027）. The results of tube formation assay showed that the number of tubule formation of HUVEC in B7-H3^＋ TEMs group（55.25±11.48） was significantly greater than that of B7-H3^-TEMs group （31.34±8.45）and blank control group（25.00±6.74） （P〈0.001）. The results of mouse aortic ring assay showed that the number of neovascularization in B7-H3^＋ TEMs group（77.35±18.47） was significantly greater than that of B7-H3^-TEMs group（39.42±8.29） and blank control group（28.79±7.63） （P〈0.001）.ConclusionsB7-H3^＋ TEMs can promote angiogenesis in ccRCC, and might act as an effective target in anti-angiogenic therapy for ccRCC.

关 键 词：肾透明细胞癌 B7-H3分子 Tie2表达单核巨噬细胞 微血管密度 血管生成 

分 类 号：R737.11[医药卫生—肿瘤]