siRNA干扰MMP-9和FAK双基因抑制小鼠黑色素瘤生长和在体迁移

目的:观察干扰MMP-9和FAK双基因对恶性黑色素瘤高转移细胞B16F10体内转移的影响。方法:构建PGV102-MMP9-siRNA、PGV102-FAK-siRNA重组质粒载体,脂质体^TM2000介导转染小鼠黑色素瘤B16F10细胞,RT-PCR检测基因的干扰效果;建立C57BL/6小鼠皮下移植瘤模型观察细胞在体成瘤和肿瘤的生长情况,常规组织切片,H&E染色观察肿瘤组织病理学特征;经C57BL/6小鼠尾静脉注射细胞5×10⁵个/只,24天后计数小鼠肺转移结节数评价肿瘤细胞在体迁移能力。结果:RT-PCR结果表明,重组质粒转染细胞组的MMP-9和FAK的mRNA水平显著低于正常细胞组(P<0.01),转染细胞组C57BL/6小鼠皮下成瘤的肿瘤生长速率、黑色素瘤肺转移结节数明显低于正常细胞组(P<0.01)。结论:干扰B16F10细胞MMP-9和FAK双基因可明显抑制小鼠体内恶性肿瘤的生长和迁移。

siRNA Inhibits the Growth and Migration of Mouse Melanoma by MMP-9 and FAK Gene

Objective:To observe the effect of combined-silencing MMP-9 and FAK on migration of malignant melanoma highly metastatic B16F10 cells in vivo. Methods:Construct recombinant plasmid vectors pGV102-MMP9-siRNA and pGV102-FAK-siRNA, then Lipofectamine ^TM2000 mediated the plasmid vectors were transfected into B16F10 cells. To observe the growth of the tumor, the C57BL/6 mouse model of subcutaneous transplantation was established, and the pathological characteristics of tumor tissue was observed by H&E staining. B16F10 cells were stably intravenously injected into C57BL/6 mice by 5×10⁵. After 24 days of injection, the number of mouse lung metastasis nodules was counted to evaluate the migration ability of tumor cells in vivo. Results:RT-PCR results showed that the mRNA level of MMP-9 and FAK in the transfected cells was significantly reduced than that of normal cells(P<0.01). Compared with the normal cells, the tumor growth rate and the number of lung metastasis nodules of C57BL/6 mice in the transfected cells were significantly reduced(P<0.01). Conclusion:Interference MMP-9 and FAK can significantly inhibit malignant tumor growth and metastasis in vivo.