重组Ⅱ型腺相关病毒介导的 TIMP-1感染 B16细胞对其体外侵袭及体内成瘤性的抑制作用研究

摘要

Objective To investigate the effect of recombinant adenovirus mediated TIMP-1 on the invasion of B16 cell in vitro, and the mechanism of tumorigenicity in vivo. Method Reconstructed plasmids that carries TIMP1 (rAAV2-TIMP1) were prepared, and B16 cells were cultured and infected with rAAV2-TIMP1 in vitro, and the expression of TIMP-1 in B16 cells were examined using Western blotting assay. Then B16 cells that expressed TIMP-1 were either injected into the subcutaneous tissue or caudal vein of the mice, and the effect of TIMP-1 on tumor formation was investigated. Result TIMP-1 was expressed effectively in B16 cells. The result showed that the TIMP-1 mediated by rAAV2 could significantly inhibit the invasion of B16 cells in vitro, and could significantly inhibit the tumorigenicity of B16. Conclusion TIMP-1 mediated by rAAV2 can inhibit the invasion of B16 in vitro, and can significantly inhibit its tumorigenicity in vivo.%目的：研究重组Ⅱ型腺相关病毒（AAV2）介导的 TIMP-1对小鼠恶性黑色素瘤细胞（B16细胞）侵袭性及体内成瘤性的抑制作用。方法重组构建携带金属蛋白酶组织抑制因子-1（TIMP-1）的 AAV2（rAAV2-TIMP1）质粒，以 rAAV2-TIMP1体外感染 B16细胞，经蛋白质印迹法（Western blotting）检测 TIMP-1表达。选择能够明确表达 TIMP-1的 B16细胞，并根据其对肿瘤细胞侵袭性的抑制作用探究 TIMP-1的作用，将感染 rAAV2-TIMP1的 B16细胞接种至小鼠的皮下与尾静脉，进而观察 AAV2介导的 TIMP-1对体内成瘤情况的影响。结果 TIMP-1经 AAV2成功导入 B16细胞并获得有效表达，表明 AAV2介导的 TIMP1能够明显抑制体外肿瘤细胞的侵袭性，明显降低体内肿瘤细胞的成瘤性。结论 AAV2介导的 TIMP1在体外能很好地抑制 B16细胞的侵袭性，在体内可以明显抑制肿瘤的形成和生长。