sFlt-1基因阻断VEGF抑制小鼠骨肉瘤生长的机制研究  

Study on the mechanism of sFlt-1 gene blocking VEGF inhibits the growth of osteosarcoma in mice

作　　者：宋文真 杨福俊[2] 李宏达 王鹏[3] 杨上游 殷德振[3] SONG Wenzhen;YANG Fujun;LI Hongda;WANG peng;YIN Dezhen(Second Clinical Medical School of Binzhou Medical University,Yantai,Shandong 264033,China;Weihai Municipal Hospital,Weihai,Shandong 264200,China)

机构地区：[1]滨州医学院第二临床医学院,山东烟台264033 [2]威海市立医院肿瘤科,山东威海264200 [3]威海市立医院脊柱骨科,山东威海264200 [4]堪萨斯大学威奇塔医学院骨外科,美国堪萨斯州威奇塔市67214

出　　处：《中华肿瘤防治杂志》2024年第7期414-419,共6页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的探讨可溶性血管内皮生长因子(VEGF)受体Flt-1(sFlt-1)与小鼠骨肉瘤生长的关系及其相关分子生物学机制。方法通过逆转录病毒载体将sFlt-1和LacZ基因转导至人骨肉瘤G-292细胞,将其与未转导的G-292细胞分别移植到重度联合免疫缺陷小鼠胫骨近端建立骨肉瘤模型,实验分为G-292组、LacZ转导组、sFlt-1转导组和假手术组。第2、4、6和8周使用microCT监测肿瘤的发展,并在8周时收集肿瘤标本检测VEGF、sFlt-1、CD34、Ki-67、c-myc、c-fos、趋化因子受体4(CXCR4)表达;实时荧光定量聚合酶链式反应(qRT-PCR)检测c-myc和c-fos mRNA表达。结果8周时sFlt-1转导组肿瘤大小为(7.56±2.00)mm^(3),小于LacZ转导组〔(28.28±6.85)mm^(3)〕和G-292组〔(32.76±10.06)mm^(3)〕,F=17.789,P<0.001。肿瘤组织中sFlt-1转导组CD34阳性微血管密度为13.60±3.27,低于LacZ转导组(24.50±6.88)和G-292组(22.50±4.74),F=12.532,P<0.001。sFlt-1转导组c-myc水平为399.35±72.14,低于LacZ转导组(672.35±108.74)及G-292组(640.11±91.43),F=115.623,P<0.001;sFlt-1转导组Ki-67水平为416.35±37.92,低于LacZ转导组(872.25±87.75)及G-292组(896.15±80.61),F=412.285,P<0.001;sFlt-1转导组CXCR4水平为400.57±13.38,低于LacZ转导组(1024.75±63.60)及G-292组(960.34±60.20),F=1033.337,P<0.001。sFlt-1转导组c-myc mRNA表达水平为5.42±0.74,低于LacZ转导组(7.60±1.14)及G-292组(8.06±1.26),F=17.380,P<0.001;c-fos mRNA表达水平为3.96±1.09,低于LacZ转导组(4.83±1.11)及G-292组(4.73±0.96),F=2.034,P=0.150。结论sFlt-1基因转导抑制了小鼠骨肉瘤的生长。sFlt-1过表达可能通过抑制肿瘤内微血管发育和下调c-myc、Ki-67和CXCR4表达抑制VEGF功能。Objective To explore the relationship between the soluble vegf receptor Flt-1(sFlt-1)and osteosarcoma growth and its possible molecular mechanism on a mouse osteosarcoma model.Methods sFlt-l and LacZ genes were transduced into human osteosarcoma G-292 cells by retroviral vectors,and the transduced and untransduced G-292 cells were respectively transplanted into the proximal tibia of severe combined immunodeficiency mice to establish osteosarcoma models.The experiment was divided into G-292 group,LacZ transduction group,sFlt-1 transduction group and sham surgery group.The tumors were monitored by microCT at 2,4,6 and 8 weeks,and tumor specimens were collected at 8 weeks for detection of VEGF,sFlt-1,CD34,Ki67,cmyc,cfos,chemokine receptor 4(CXCR4)expression,and real-time fluorescence quantitative(qRT)-PCR for detection of cmyc and cfos mRNA expression.Results At 8 weeks,the tumor size in sFlt-1 transduction group was(7.56±2.00)mm^(3),which was smaller than that in LacZ transduction group[(28.28±6.85)mm^(3)] and G-292 group[(32.76±10.06)mm^(3)],F=17.789,P<0.001.The CD34-positive microvascular density in sFlt-1 transduction group was 13.60±3.27,which was lower than that in LacZ transduction group(24.50±6.88)and G-292 group(22.50±4.74),F=12.532,P<0.001.The level of cmyc in sFlt-1 transduction group was 399.35±72.14,lower than that in LacZ transduction group(672.35±108.74)and G-292 group(640.11±91.43),F=115.623,P<0.001.Ki-67 level in sFlt-1 transduction group was 416.35±37.92,lower than that in LacZ transduction group(872.25±87.75)and G-292 group(896.15±80.61),F=412.285,P<0.001.The CXCR4 level in sFlt-1 transduction group was 400.57±13.38,lower than that in LacZ transduction group(1024.75±63.60)and G-292 group(960.34±60.20),F=1033.337,P<0.001.The expression level of c-myc mRNA in sFlt-1 transduction group was 5.42±0.74,lower than that in LacZ transduction group(7.60±1.14)and G-292 group(8.06±1.26),F=17.380,P<0.001.The mRNA expression level of c-fos was 3.96±1.09,which was lower than that of LacZ trans

关 键 词：骨肉瘤 可溶性血管内皮生长因子受体 血管内皮生长因子 微血管密度 

分 类 号：R738.1[医药卫生—肿瘤]