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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 |
谢群1△,唐南洪2,3△,林扬元1,林建银2* |
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(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 △共同第一作者 *通信作者) |
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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。 |
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 |
DOI:10.3724/SP.J.1008.2014.00668 |
投稿时间:2013-09-19修订日期:2014-02-26 |
基金项目: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). |
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Effect of recombinant adenovirus harboring snake venom cystatin on invasion and metastasis of human hepatocellular carcinoma cells MHCC97H |
XIE Qun1△,TANG Nan-hong2,3△,LIN Yang-yuan1,LIN Jian-yin2* |
(1. School of Basic Medical Science, Putian University, Putian 351100, Fujian, China; 2. Research Center of Molecular Medicine, Key Laboratory of Ministry of Education for Gastrointestinal Cancer, Fujian Medical University, Fuzhou 350004, Fujian, China; 3. Hepatobiliary Surgery Institute, Union Hospital, Fujian Medical University, Fuzhou 350001, Fujian, China △Co-first authors. *Corresponding authors.) |
Abstract: |
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. |
Key words: snake venom cystatin adenovirus hepatocellular carcinoma neoplasm invasiveness neoplasm metastasis |