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目的观察胃癌细胞系SGC7901转染Twist基因后细胞中AP-1转录活性及基质金属蛋白酶-9(MMP-9)表达的改变,探讨Twist促进肿瘤侵袭转移的可能机制。方法利用DNA的限制性内切酶技术鉴定质粒PcDNA3-Twist;采用脂质体法将Twist基因稳定转染胃癌细胞系SGC7901,应用Western-blot检测Twist表达;电泳迁移率变动分析(EMSA)测定转染前后AP-1的DNA结合活性;Western-blot检测转染前后AP-1下游分子MMP-9的表达。结果SGC7901细胞转染质粒Twist-PcDNA3后Twist表达明显上升,AP-1的DNA结合活性明显增强。同时,转染阳性质粒Twist-PcDNA3后细胞的MMP-9表达明显上升。结论Twist促进肿瘤侵袭转移可能存在Twist/AP-1/MMP-9介导的通路。
Objective To observe the changes of AP-1 transcriptional activity and the expression of matrix metalloproteinase-9 (MMP-9) in human gastric cancer cell line SGC7901 transfected with Twist gene and to explore the possible mechanism by which Twist promotes tumor invasion and metastasis. Methods The plasmid pcDNA3-Twist was identified by DNA restriction endonuclease assay. Twist gene was stably transfected into gastric cancer cell line SGC7901 by lipofectamine and the expression of Twist was detected by Western-blot. The electrophoretic mobility shift assay (EMSA) DNA-binding activity of AP-1 before and after dyeing; Western blot was used to detect the expression of MMP-9 downstream of AP-1 before and after transfection. Results Twist-PcDNA3 transfected SGC7901 cells significantly increased Twist expression, AP-1 DNA binding activity was significantly enhanced. At the same time, the expression of MMP-9 in transfection of positive plasmid Twist-PcDNA3 was significantly increased. Conclusion Twist / AP-1 / MMP-9-mediated pathway may be involved in the invasion and metastasis of tumor.