ZFX基因在骨肉瘤中的表达及其作用机制

Expression and Mechanism of ZFX Gene in the Osteosarcoma Cells

目的探讨X染色体偶联锌指蛋白(ZFX)基因在骨肉瘤中的表达及其作用机制。方法采用手术切除后存档的人骨肉瘤组织及瘤旁组织石蜡包埋标本50例,通过Western blot检测其中ZFX的表达。体外培养人骨肉瘤MG63细胞,分别用ZFX小干扰RNA(ZFX-siRNA)和阴性对照(siRNA-NC)转染细胞,以空脂质体转染的细胞作为对照组,各组细胞培养48 h后,用Western blot检测细胞中ZFX、Bcl-2、Bax、β-catenin、Cyclin D1蛋白表达变化;CCK-8检测细胞增殖;流式细胞术检测细胞凋亡。结果 ZFX基因在骨肉瘤组织中的蛋白表达显著高于瘤旁组织(P<0.01);siRNA-NC组细胞增殖活力、凋亡率及ZFX、Bcl-2、Bax、β-catenin、Cyclin D1蛋白表达与对照组比较无显著差异(P>0.05);ZFX-siRNA组细胞活力及ZFX、Bcl-2、β-catenin、Cyclin D1蛋白表达显著低于对照组,细胞凋亡率及Bax蛋白表达显著高于对照组(P<0.01)。结论 ZFX基因在人骨肉瘤组织中高表达,可能通过促进人骨肉瘤MG63细胞增殖并抑制其凋亡在其发生发展过程中发挥重要的作用,其机制可能与Wnt信号通路的激活有关。

Objective To investigate the expression and mechanism of zinc finger protein X-linked （ZFX） gene in the osteosarcoma cells. Methods The paraffin embedded specimens were collected from 50 cases of human osteosarcoma tissues and adjacent tissues after surgi- cal resection. The expressions of ZFX in the specimens were detected by Western blot. Human osteosarcoma MG63 cells were cultured in vitro, and ZFX small interfering RNA （ZFX-siRNA） and negative control （siRNA-NC） were respectively transfected into cells. The empty liposomes transfected cells were taken as the control group. After the cells were cultured for 48h, the protein expressions of ZFX, Bcl-2, Bax,β-catenin and CyclinD1 were detected by Western blot. Cell proliferation activity was detected by CCK-8, and cell apoptosis rate was determined by flow cytometry. Results The protein expression of ZFX gene in osteosarcoma tissues was significantly higher than in tumor adjacent tissues（P〈 0.01）. No significant differences were found between the siRNA-NC group and the control group in cell proliferation ac- tivity, apoptosis rate and protein expressions of ZFX, Bcl-2, Bax,β-catenin and CyclinD 1 （P 〉 0.05）. However, the cells proliferation activ- ity and protein expressions of ZFX, Bcl-2,β-catenin and CyclinD1 in ZFX-siRNA group were significantly lower than in control group. The cell apoptosis rate and expression of Bax protein were significantly higher in ZFX-siRNA group than in control group （P 〈 0.01）. Conclusion The ZFX gene was highly expressed in human osteosarcoma tissues. It may play an important role in promoting the proliferation and inhibiting the apoptosis of human osteosarcoma MG63 ceils. The mechanism may be related to the activation of Writ signaling pathway.