髓锌指基因1对骨肉瘤细胞增殖和凋亡的影响

Effect of MZF -1 on proliferation and apoptosis osteosarcoma cells

摘要:

目的研究葡萄糖-6- 磷酸脱氢酶（G6PD）和髓锌指基因1（MZF -1）在骨肉瘤中的作用及其可能的作用机制。方法实时荧光定量聚合酶链反应（qRT-PCR）检测MZF -1 和G6PD在骨肉瘤细胞中的表达；MTT 法检测过表达MZF -1 对骨肉瘤细胞增殖的影响；流式细胞仪检测过表达MZF -1 对骨肉瘤细胞凋亡及细胞周期的影响；荧光素酶报告基因分析法验证MZF-1 与G6PD启动子区的相互作用关系。结果在骨肉瘤细胞中， MZF -1 的表达下降，G6PD表达上升。过表达MZF -1可抑制骨肉瘤细胞增殖，促进细胞凋亡，并且将细胞周期阻滞于G0/G1期。荧光素酶报告基因分析结果表明， MZF -1可靶向结合到G6PD 的启动子区域并负向调控G6PD 的表达。结论MZF -1 可通过负向调节G6PD的表达，在骨肉瘤的发生、发展过程中发挥抑癌的作用，为骨肉瘤治疗提供新的思路。

Abstract:

Objective To investigate the effect of myeloid zinc finger gene 1 (MZF -1 ) and glucose-6- phosphate dehydrogenase (G6PD) on progression of osteosarcoma and the underlying mechanisms. Methods The expression levels of MZF -1 and G6PD were determined by real-time PCR. MTT was utilized to examine the cellular proliferation. Apoptosis rate and cell cycle were measured by flow cytometry. The molecular interaction between MZF -1 and G6PD was manifested by dual-luciferase reporter assay. Results MZF -1 was down -regulated and G6PD was up -regulated in osteosarcoma cells. Over-expression of MZF -1 inhibited osteosarcoma cell proliferation, increased cellular apoptosis, and induced cell cycle arrest on G0/G1 phase.Dual-luciferase reporter assay showed that MZF -1 bonded to the promoter region of G6PD and negatively regulated the transcription of G6PD. Conclusions MZF -1 exerts as a tumor suppressor by negatively regulating the expression of G6PD during the progression of osteosarcoma.