Study on the mechanism of sFlt-1 gene blocking VEGF inhibits the growth of osteosarcoma in mice
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-1 and LacZ genes were transduced into human osteosarcoma G-292 cells by retroviral vectors,and the transduced and untransduced G-292 cells were respec-tively 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,Ki-67,c-myc,c-fos,chemokine receptor 4(CXCR4)expression,and real-time fluorescence quantitative(qRT)-PCR for detection of c-myc and c-fos mRNA expression.Results At 8 weeks,the tumor size in sFlt-1 transduction group was(7.56±2.00)mm3,which was smaller than that in LacZ transduction group[(28.28± 6.85)mm3]and G-292 group[(32.76±10.06)mm3],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 c-myc 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=1 033.337,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 transduction group(4.83±1.11)and G-292 group(4.73±0.96),F=2.034,P=0.150.Conclusions sFlt-1 gene transduction inhibits the growth of os-teosarcoma in mice.Overexpression of sFlt-1 may inhibit VEGF function by inhibiting microvascular development in tumors and down-regulating the expression of c-myc,Ki-67 and CXCR4.
osteosarcomasoluble vascular endothelial growth factor receptorvascular endothelial growth factormicro-vascular density
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