Runt相关转录因子3对人骨肉瘤细胞促血管形成能力的影响

Effect of Runt-related transcription factor-3 on angiogenesis abilities of human osteosarcoma cells

目的研究Runt相关转录因子3(RUNX3)对人骨肉瘤细胞株U2OS促血管形成能力的影响。方法 U2OS细胞转染RUNX3过表达质粒,免疫印迹(Western blot)法检测细胞内血管内皮生长因子(VEGF)、黏着斑激酶(FAK)蛋白表达变化,酶联免疫吸附(ELISA)法测定VEGF水平的变化。细胞计数(CCK-8)法检测RUNX3过表达对人脐静脉内皮细胞(HUVECs)增殖的影响,微血管形成实验测定RUNX3过表达对HUVECs微血管形成的影响。结果转染RUNX3质粒后,实验组U2OS细胞中RUNX3基因为2.20±0.22,RUNX蛋白0.25±0.04,对照组RUNX3基因为1.40±0.11,RUNX3蛋白为0.15±0.03,差异有统计学意义(P<0.05)。RUNX3过表达后,U2OS细胞中FAK和VEGF蛋白分别为0.15±0.02和0.22±0.03,对照组分别为0.37±0.05,和0.42±0.05,差异有统计学意义(P<0.05);实验组VEGF水平为(2381.04±166.07)pg·m L^(-1),对照组为(3177.74±322.76)pg·m L^(-1),差异有统计学意义(P<0.05)。RUNX3过表达后,U2OS细胞的促HUVECs血管形成个数为(5.00±1.00)个,对照组为(12.00±2.00)个;实验组U2OS促HUVECs增殖活性为0.66±0.02,对照组为1.05±0.09,差异有统计学意义(P<0.05)。结论 RUNX3可抑制骨肉瘤细胞的促血管形成能力,其机制可能是通过减少细胞中FAK的表达、抑制细胞对VEGF的分泌来实现的。

Objective To investigate the effect of Runt- related tran- scription factor -3 （RUNX3） on angiogenesis ability of human osteosarcoma U2OS cells. Methods RUNX3 over- expression plasmid was transfected to increase RUNX3 expression, then cells were subjected to cell counting kit （CCK- 8 ） assay, tube formation assay to detect RUNX3 over- expression on cell angiogenesis ability. Western blot and enzyme- linked immunosorbent assay （ELISA） were used to detect vas- cular endothelial growth factor （VEGF） and focal adhension kinase （FAK） expression and activities. Results After transfection, R UNX3 gene and RUNX3 protein in test group were 2.20 ± 0.22, 0. 25 ± 0. 04, had significant difference with control group, which were 1.40 ± 0. 11, 0. 15 ±0. 03 （P 〈 0.05）. After transfection, FAK and VEGF protein in test group were 0. 15 ± 0. 02, 0.22 ± 0. 03, had significant difference with control group, which were 0. 37 ± 0. 05 and 0.42 ± 0.05 （P 〈 0. 05 ）.The VEGF in test and control group were （2381.04 ± 166.07 ）, （ 3177.74 ± 322. 76 ） pg . mL-1 （ p 〈 0. 05 ）. The number of tube formation after forced expression of RUNX3 was 5.00 ± 1.00, had significant difference with 12.00 ± 2.00 in control group （P 〈 0. 05 ）. The cell proliferation activity of HUVECs in test group was 0. 66 ± 0. 02, had significant difference with 1.05 ± 0. 09 in control group （P 〈 0. 05 ）. Conclusion Over - expression of RUNX3 results in the inhibition of cell angiogenesis abilities of human osteosarcoma U2OS cells.