脂肪细胞谷氨酰胺合成酶抑制结肠癌细胞的腹腔转移及血管生成  

Glutamine synthetase from adipocytes inhibits peritoneal metastasis and angiogenesis of colon cancer in mice

作　　者：陈晓珍 李一飞 张璇[1] 缪洪明[1] 梁后杰[1] 

机构地区：[1]第三军医大学西南医院肿瘤科,重庆400038

出　　处：《第三军医大学学报》2018年第1期70-75,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81702929)~~

摘　　要：目的探讨脂肪细胞谷氨酰胺合成酶(glutamine synthetase,GS)对结肠癌腹腔转移及血管生成的影响。方法转基因小鼠的鉴定:Western blot检测野生型(WT)小鼠及脂肪细胞GS转基因(TgGS)小鼠脂肪细胞GS蛋白的表达。体内实验:分别构建WT小鼠及TgGS小鼠的腹腔转移模型,观察结肠癌细胞在腹腔的种植情况;免疫组化检测移植瘤的血管生成情况。体外实验:取WT及TgGS小鼠的脂肪进行无菌培养,收集条件培养基;通过小管形成实验、CCK8实验、Transwell实验观察条件培养基对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)的小管形成、增殖及迁移的影响。结果 Western blot检测结果显示,TgGS小鼠较WT小鼠脂肪细胞GS蛋白水平显著上调(P<0.05)。TgGS小鼠较WT小鼠腹腔移植瘤数量明显减少[(6.33±4.04)vs(16.00±6.56),P<0.05],体积明显减小[(2.35±1.93)vs(9.37±5.07),P<0.01]。免疫组化结果显示,TgGS小鼠肿瘤血管生成明显少于WT小鼠(P<0.05)。体外小管形成实验显示TgGS组较WT组HUVECs小管形成总长度明显缩短(P<0.05);CCK8实验显示TgGS组及WT组HUVECs的增殖差异无统计学意义(P>0.05);Transwell实验显示TgGS组HUVECs细胞迁移数量明显少于WT组[(15.33±5.50)vs(78.33±13.58),P<0.01]。结论脂肪细胞GS可抑制结肠癌腹腔转移及肿瘤血管生成。Objective To determine the effect of glutamine synthetase （GS） derived from adipocytes on intraperitoneal metastasis and angiogenesis of colon cancer. Methods Wide type（WT）and transgenetic mice （TgGS） were firstly identified with Western blotting for the expression level of GS protein in the adipocytes.Then mouse model of peritoneal metastasis of colon cancer MC38 cells was established by intraperitoneally injecting 2.0×106 cells in the WT mice and TgGS mice. Then the cell implantation and tumor formation were observed.Immunohistochemical assay was used to detect the angiogenesis of the tumor mass.The fat tissues were harvested from WT and TgGS mice,and then the adipocytes were isolated and cultured to obtain the 2 kinds of adipocyte conditional media.Then tube formation assay, CCK8 assay, and Transwell chamber test were used to determine the effects of the 2 kinds of conditioned media on the tube formation, proliferation and migration in human umbilical vein endothelial cells （HUVECs）. Results Western blotting showed that the expression level of GS protein in the adipocytes of TgGS mice was significantly increased than that in WT mice （P〈0.05）. In the xenograft model, there were notably less （6.33±4.04 vs 16.00±6.56,P〈0.05） and smaller（2.35±1.93 vs 9.37±5.07, P〈0.01） tumors in the TgGS mice when compared with the WT mice. Immunohistochemical assay showed that the angiogenesis in TgGS mice were obviously less than that in WT mice（P〈0.05）.The tube formation assay indicated that the total lengths of tubules was significantly shorter in the HUVECs co-cultured with adipocyte conditional medium from the TgGSmice than those with the medium from the WT mice group （P〈0.05）. CCK8 assay displayed that there was no significant difference in the cell proliferation of HUVECs between the 2 kinds of media （P〉0.05）.Transwell chamber test showed that there were less migrated cells in the HUVECs with TgGS culture medium than those with WT culture medium （15.33±5

关 键 词：脂肪细胞谷氨酰胺合成酶 血管生成 结肠癌 腹腔转移 

分 类 号：R345[医药卫生—基础医学] R730-37