摘要
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。
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） MHCC97 H 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 szrcystatin 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.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2014年第6期668-671,共4页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(30371747)
福建省教育厅资助省属高校科研专项(JK2011050)
莆田市科技计划项目[2012S11(1)]
福建省自然科学基金指导性科技计划项目(2012D117)~~
