沉默Runt相关转录因子3对骨肉瘤细胞恶性程度的影响

Effect of silencing Runt-related transcription factor 3 on malignancy of osteosarcoma cells

目的探讨沉默Runt相关转录因子3(RUNX3)对骨肉瘤细胞增殖能力及血管新生能力的影响。方法培养骨肉瘤HEK293细胞株并将其分为NC组和RUNX3组,分别转染阴性对照小干扰RNA(siRNA)和RUNX3-siRNA。于转染siRNA后6、12、24、48 h采用MTS试剂盒测定细胞增殖活力;转染siRNA后24、48 h采用荧光定量PCR试剂盒测定细胞中增殖分子β-链蛋白、细胞周期蛋白D1(Cyclin D1)、增殖细胞核抗原(PCNA)的mRNA表达量,采用ELISA试剂盒测定细胞培养基中血管内皮生长因子(VEGF)、成纤维细胞生长因子(b FGF)、促血管生成素-2(Ang-2)的含量。结果转染siRNA后6、12、24、48 h RUNX3组的细胞增殖活力值均高于NC组(0.78±0.11vs.0.57±0.09,0.99±0.14 vs.0.73±0.10,1.31±0.18 vs.0.81±0.12,1.72±0.26 vs.0.98±0.14,P均<0.05);转染siRNA后24、48 h RUNX3组细胞中β-链蛋白、Cyclin D1、PCNA的mRNA表达量以及细胞培养基中VEGF、b FGF、Ang-2的含量均高于NC组(P均<0.05)。结论沉默RUNX3能够增强骨肉瘤细胞的增殖能力及血管新生能力。

Objective To investigate the effect of silencing Runt-related transcription factor 3 （RUNX3） on the proliferation and angiogenesis of osteosarcoma cells. Methods Osteosarcoma cell line HEK293 was divided into the NC group （transfected with negative control-siRNA） and RUNX3 group （trans-fected with RUNX3-siRNA）. At 6h, 12 h, 24 h and 48 h after transfection with siRNA, cell proliferation was determined by MTS kit. At 24 h and 48 h after transfection with siRNA, the expression levels of p-chain pro-tein, CyclinD1 and proliferating cell nu clear antigen （PCNA） mRNA were measured by fluorescent quantita-tive PCR kit. The content of vascular endothelial growth factor （ VEGF） , promote angiogenin-2 （Ang-2 ） and fibroblast growth factor （bFGF） in the culture medium was quantitatively determined by enzyme-linked immu-nosorbent assay kit. Results At 6 h , 12 h , 24 h and 48 h after transfection with siRNA, the activity of cell proliferation in the RUNX3 group was significantly higher than that in the NC group （0. 78 ±0. 11 0. 57 ±0.09, 0.99 ±0. 14 vs .0.73 ±0. 10,1.31 ±0 .18 vs.0 .81 ± 0.12,1.72 ± 0.26vs. 0.98 ± 0.14 ,all P〈0.05）. At 24 h and 48 h after transfection with siRNA, the expression levels of p-chain protein, CyclinDl and PCNA mRNA in cells and VEGF, Ang-2 and bFGF in the cultured medium in the RUNX3 group were signifi-cantly higher compared with those in the NC group （all P 〈 0. 05 ）. Conclusion Silencing RUNX3 can en-hance the proliferation and angiogenesis of osteosarcoma cells.