携反义基质金属蛋白酶-2的重组腺病毒对肝癌生长的影响

Recombinant Adenovirus Vectors Carrying Antisense Matrix Metalloproteinase-2 Inhibits Hepatocellular Carcinoma Growth in vivo

目的探讨携带反义二型基质金属蛋白酶(MM P 2)基因的重组腺病毒(A d-MM P 2AS)对人肝癌细胞株(B e l-7402)体外侵袭性和人肝癌动物模型生长的抑制作用。方法用已构建的A d-MM P 2AS感染人肝癌细胞株(B e l-7402),用侵袭小室模型检测A d-MM P 2AS对B e l-7402细胞降解人工基底膜的抑制作用;用W estern B lot和明胶酶谱检测A d-MM P 2AS对B e l-7402细胞分泌MM P 2的影响;用感染了A d-MM P 2AS的B e l-7402细胞接种于裸鼠皮下,使其在裸鼠体内成瘤,以观察其成瘤能力;用A d-MM P 2AS瘤内注射,以观察它对肝癌生长的抑制作用。结果A d-MM P 2AS感染的B e l-7402细胞分泌MM P 2被抑制、穿过人工基底膜M atrige l的B e l-7402细胞数下降52%、感染A d-MM P 2AS的B e l-7402细胞的成瘤量下降4.3倍,A d-MM P 2AS瘤内注射后瘤体生长减少63%。结论A d-MM P 2AS能明显抑制肝癌的生长,对肝癌有治疗潜力。

Objective To investigate the inhibitory effect of a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2（MMP2） on the growth of hepatocellular carcinoma （HCC） in vivo. Methods The recombinant adenoviral vector carrying antisense MMP2（Ad-MMP2^AS）which had been constructed by us in readiness was used to infect the human HCC cell line （Bel-7402）. Then the invasiveness of the Bel-7402 cells was assayed in Matrigel, and the production of MMP2 in the Bel-7402 cells was detected with Western blot analysis and Gelatin zymography. After the Ad-MMP2^AS-infected Bel-7402 cells being subcutaneously inoculated in nude mice, the production of tumors was under observation, and then Ad-MMP2^AS was injected intratumorally into the pre-existing tumors. Results Compared with PBS or Ad-CMV-infected cells, infection of Bel-7402 cells with Ad-MMP2^AS significantly reduced MMP2 enzyme activity, the invasiveness resulted in 52% reduction in Matrigel assays, and the tumor volume displayed a 4. 3-fold reduction in nude mice. In addition, direct intratumoral injection of Ad-MMP2^AS into pre-existing tumors significantly impaired the further expansion of the tumor mass and resulted in a 63% reduction in tumor cell growth. Conclusion The recombinant adenovirus with antisense MMP2 can effectively inhibit the invasiveness and growth of Bel-7402 cells in vitro and in vivo, and has a therapeutic potential for HCC.