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  

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

作　　者：谢群[1] 唐南洪[2,3] 林扬元[1] 林建银[2] 

机构地区：[1]福建省莆田学院基础医学院,莆田351100 [2]福建医科大学分子医学研究中心,消化道恶性肿瘤教育部重点实验室,福州350004 [3]福建医科大学附属协和医院省肝胆外科研究所,福州350001

出　　处：《第二军医大学学报》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)~~

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.

关 键 词：蛇毒半胱氨酸蛋白酶抑制剂 腺病毒 肝细胞癌 肿瘤侵袭 肿瘤转移 

分 类 号：R735.7[医药卫生—肿瘤]