腺病毒介导反义基质金属蛋白酶-2基因抑制肝癌生长的实验研究  

Inhibition of adenovirus-mediated gene transfer of antisensc matrix metalloproteinase-2 on hepatocellular carcinoma growth in vivo

作　　者：张明满[1] 严律南[1] 李德华 苟兴华 刘江文[1] 苏芝 韩蕾 赵兰英 

机构地区：[1]四川大学华西医院普外科,成都610041 [2]成都地奥制药集团有限公司新药部

出　　处：《中华肝脏病杂志》2005年第9期671-674,共4页Chinese Journal of Hepatology

摘　　要：目的探讨携带反义基质金属蛋白酶-2(MMP2)基因的重组腺病毒(Ad-MMP2AS)对人肝癌细胞株(Bel-7402)体外侵袭性和人肝癌动物模型生长的抑制作用。方法Ad-MMP2AS感染人肝癌细胞Bel-7402后,用Boyden Chamber检测Ad-MMP2AS对Bel-7402细胞降解人工基底膜Matrigel的抑制作用; Western blot和明胶酶谱分析测定Ad-MMP2AS对Bel-7402细胞分泌MMP2的影响。将感染Ad-MMP2AS的Bel-7402细胞接种于裸鼠皮下观察其成瘤能力;Ad-MMP2AS瘤内注射,观察它对肝癌生长的抑制作用;肿瘤组织切片、HE染色观察瘤细胞生长情况。结果Ad-MMP2AS感染的Bel-7402细胞分泌MMP2被抑制、穿过Matrigel的Bel-7402细胞数下降52.05%、成瘤量下降3.3倍;瘤内注射Ad-MMP2AS后瘤体生长抑制率为63.06%。结论重组腺病毒介导的反义MMP2基因(Ad-MMP2AS)能抑制肝癌的生长,对肝癌有治疗潜力。Objective To investigate if a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2 （MMP2） gene would inhibit the growth of hepatocellular carcinoma （HCC） in vivo. Methods Using the recombinant adenoviral vector carrying antisense MMP2 gene （Ad-MMP2^AS） which was constructed by us previously, 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 blotting analysis and Gelatin zymography. Then the Ad-MMP2^AS-infected cells were subcutaneously inoculated in nude mice. After tumors developed, Ad-MMP2^AS was injected intratumorally into pre-existing tumors. The tumors were removed, sectioned, and stained with H＆E. Results Compared with PBS or Ad-CMV-infected cells, the infected Bel-7402 cells with Ad-MMP2^AS injections significantly reduced their MMP2 enzyme activity and invasiveness about 52.05% in Matrigel assays, and the tumor volumes in nude mice resulted in a 3.3-fold reduction. In addition, direct intratumoral injection of Ad-MMP2%^AS into pre-existing tumors significantly prevented further expansion of the tumor masses and resulted in a 63.06% 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 it has a therapeutic potential for HCC.

关 键 词：癌 肝细胞 基质金属蛋白酶 基因疗法 腺病毒 人 基质金属蛋白酶-2 重组腺病毒介导 人肝癌细胞株 介导反义 人肝癌细胞BEL-7402 基因抑制 

分 类 号：R735.7[医药卫生—肿瘤]