Artemin在软骨肉瘤中的表达及其对内皮细胞增殖和迁移的影响

Artemin expression in chondrosarcoma and its effect on proliferation and migration of endothelial cells

目的:探讨Artemin在软骨肉瘤中的表达水平及其对内皮细胞增殖与迁移的作用及其机制。方法:收集2015年5月至2019年4月南京中医药大学连云港附属医院手术切除的40例软骨肉瘤患者的肿瘤组织标本,分为低(Ⅰ级)、高级别(Ⅱ、Ⅲ级)软骨肉瘤各20例;对照组为20例因车祸等意外伤截肢患者的正常软骨组织标本。用免疫组化法检测肿瘤组织中Artemin、血管内皮生细胞长因子(vascular endothelial growth factor,VEGF)、Ki-67表达水平和CD31^+血管密度。用10 ng/ml的Artemin处理SW1353细胞后,用ELISA实验检测细胞培养上清中VEGF、基质细胞衍生因子-1(stromal cell derived factor-1,SDF-1)、基质金属蛋白酶2(matric metalloproteinase 2,MMP2)和MMP9的含量变化,用Transwell迁移实验和MTT细胞增殖实验分别检测Artemin处理的软骨肉瘤细胞对ECV304细胞增殖和迁移的影响。结果:低级别组软骨肉瘤组织中Artemin和Ki-67表达水平均显著高于对照组(均P<0.01),高级别组软骨肉瘤组织中Artemin表达水平和Ki-67表达率显著高于低级别组(均P<0.01)。Artemin表达与软骨肉瘤组织中VEGF水平和血管密度呈正相关(均P<0.01);Artemin促进软骨肉瘤细胞分泌VEGF,而对SDF-1、MMP2和MMP9分泌无显著的影响;Artemin通过促进软骨肉瘤细胞分泌VEGF,诱导ECV304细胞增殖和迁移(均P<0.01)。结论:Artemin在软骨肉瘤组织中高表达且与VEGF水平和血管密度呈正相关,Artemin能够增强软骨肉瘤细胞诱导血管生成功能。

Objective:To investigate the expression of Artemin in chondrosarcoma and its effect on proliferation and migration of endothelial cells,and to explore the mechanism.Methods:A total of 40 chondrosarcoma tissue samples(low degree(Ⅰ),20 cases;high degree(Ⅱ,Ⅲ),20 cases)surgically resected from patients,who were treated in Lianyungang Hospital Affiliated to Nanjing University of Chinese Medicine from May,2015 to April,2019,were collected for this study.Another 20 cases of normal cartilage tissue specimen from patients with amputations due to car accidents were served as control.The expressions of Artemin,vascular endothelial growth factor(VEGF),Ki-67 and CD31^+vascular density in tumor tissues were detected by immunohistochemistry.After being treated with 10 ng/ml Artemin,the changes of VEGF,stromal cell derived factor-1(SDF-1),matric metalloproteinase 2(MMP2)and MMP9 in the supernatant of SW1353 cell culture were detected by enzyme-linked immunosorbent assay(ELISA),and the effects of Artemin-treated chondrosarcoma cells on the migration and proliferation of ECV304 cells were detected by Transwell migration assay and MTT cell proliferation assay,respectively.Results:The expressions of Artemin and Ki-67 in the tissues of low-level group were significantly higher than those in the control group(all P<0.01);the expressions of Artemin and Ki-67 in the tissues of high-level group were significantly higher than those in the low-level group(all P<0.01).The expression of Artemin was positively correlated with VEGF level and vascular density in chondrosarcoma tissues(all P<0.01);Artemin promoted the secretion of VEGF by chondrosarcoma cells,but had no significant effect on the secretion of SDF-1,MMP2 and MMP9.Artemin induced the proliferation and migration of ECV304 cells by promoting the secretion of VEGF by chondrosarcoma cells(all P<0.01).Conclusion:Artemin is highly expressed in chondrosarcoma tissues and has a positive correlation with the expression of VEGF and vascular density.Artemin can enhance the angiogenesis induced by chondrosarcoma.