蛇毒半胱氨酸蛋白酶抑制剂重组腺病毒对人肝癌细胞MHCC97H侵袭转移的影响

目的 构建携带蛇毒半胱氨酸蛋白酶抑制剂（sv-cystatin）基因的重组腺病毒载体（Ad/sv-cystatin），研究其对人肝癌细胞MHCC97H在体内、外侵袭转移的影响。 方法 构建携带sv-cystatin的重组腺病毒，体外感染MHCC97H细胞，采用CCK-8法检测细胞生长，Transwell小室检测细胞侵袭和迁移能力，通过裸鼠皮下肝癌细胞肺转移模型观察Ad/sv-cystatin瘤内注射对裸鼠的治疗作用。 结果 体外实验证实Ad/sv-cystatin能够感染MHCC97H细胞。与对照组及空载体组相比，Ad/sv-cystatin对MHCC97H细胞体外生长、侵袭和迁移均具有明显的抑制作用，瘤内注射Ad/sv-cystatin可以显著降低MHCC97H细胞的肺转移。 结论 重组腺病毒Ad/sv-cystatin具有抑制人肝癌细胞MHCC97H体内外侵袭转移的作用，在肝癌基因治疗方面具有开发应用潜能。

Effect of recombinant adenovirus harboring snake venom cystatin on invasion and metastasis of human hepatocellular carcinoma cells MHCC97H

Objective To construct recombinant adenovirus harboring snake venom cystatin (Ad/sv-cystatin) and to investigate its effect on invasion and metastasis (in vivo and in vitro) of human hepatocellular carcinoma (HCC) MHCC97H cells. Methods The recombinant Ad/sv-cystatin harboring sv-cystatin was constructed to infect MHCC97H cells. Then the growth of MHCC97H cells was assessed by CCK-8. Transwell matrigel assay was used to assess MHCC97H cell migration and invasiveness in vitro. Spontaneous lung metastasis assays were used to examine the effects of Ad/sv-cystatin on the invasion and metastasis of MHCC97H cells in vivo. Results Recombinant Ad/sv-cystatin harboring sv-cystatin gene could infect MHCC97H cells. Ad/sv-cystatin significantly inhibited the growth, migration and invasion of MHCC97H cells in vitro compared with control and Ad/null groups. Intra-tumoral injection of Ad/sv-cystatin significantly inhibited the lung metastasis of MHCC97H cells in nude mice compared with control and Ad/null-treated mice. Conclusion It is indicated that recombinant Ad/sv-cystatin can suppress MHCC97H cell growth, invasion and metastasis in vitro and in vivo, showing a potential for gene therapy of HCC.