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目的 :观察三氧化二砷(arsenic trioxide,As2O3)联合3’-叠氮-3’-脱氧胸腺嘧啶核苷(3’-azido-3’-deoxythymidine,AZT)对人肝癌Hep G2细胞迁移和侵袭的影响,并探讨其可能的作用机制。方法 :分别用As2O3、AZT以及两药联合处理肝癌Hep G2细胞,同时以未经药物处理的Hep G2细胞作为空白对照。应用划痕愈合实验和Transwell迁移及侵袭实验分别检测细胞迁移及侵袭能力的变化,实时荧光定量PCR法检测细胞中基质金属蛋白酶2(matrix metallopeptidase 2,MMP2)和血管内皮生长因子(vascular endothelial growth factor,VEGF)m RNA表达的变化,蛋白质印迹法检测MMP2、VEGF、细胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2,ERK1/2)和磷酸化ERK1/2(phosphorylated ERK1/2,p-ERK1/2)蛋白表达的变化。结果 :经As2O3联合AZT干预后,Hep G2细胞的迁移和侵袭能力均比空白对照及单药组明显降低(P值均<0.01)。As2O3联合AZT干预组Hep G2细胞中,MMP2和VEGF m RNA的表达水平明显低于空白对照组和各单药组(P值均<0.01),MMP2、VEGF和p-ERK1/2蛋白的表达水平也明显降低(P值均<0.01),而ERK1/2蛋白表达无明显变化(P>0.05)。结论 :As2O3联合AZT作用能够明显抑制人肝癌Hep G2细胞的迁移和侵袭能力,该抑制作用可能是通过调控ERK1/2信号通路的磷酸化以及下调MMP2和VEGF的表达来实现的。
OBJECTIVE: To observe the effect of arsenic trioxide (As2O3) and 3’-azido-3’-deoxythymidine (AZT) on the migration and invasion of Hep G2 cells, And explore its possible mechanism of action. Methods: Hep G2 cells were treated with As2O3, AZT and two drugs, respectively. At the same time, untreated Hep G2 cells were used as blank control. The changes of cell migration and invasion were detected by scratch healing assay and Transwell migration and invasion assay, respectively. The expression of MMP-2 and VEGF was detected by real-time fluorescence quantitative PCR. , VEGF) m RNA were detected by Western blotting. MMP2, VEGF, extracellular signal-regulated kinase 1/2 (ERK1 / 2) and phosphorylated ERK1 / 2 , p-ERK1 / 2) protein expression changes. Results: After As2O3 combined with AZT, the migration and invasion ability of Hep G2 cells were significantly lower than those of blank control group and single drug group (all P <0.01). The expression levels of MMP2 and VEGF m RNA in Hep G2 cells treated with As2O3 combined with AZT were significantly lower than those in blank control group and each single drug group (P <0.01), and the expressions of MMP2, VEGF and p-ERK1 / 2 (P <0.01), while the expression of ERK1 / 2 did not change significantly (P> 0.05). CONCLUSION: As2O3 combined with AZT can significantly inhibit the migration and invasion of Hep G2 cells, which may be through the regulation of phosphorylation of ERK1 / 2 signaling pathway and the down-regulation of MMP2 and VEGF expression.