摘要
目的 提高原位转基因效率 ,建立高效自杀基因杀伤胃癌肿瘤的方法。方法 首先构建含胞嘧啶脱氨酶 (CD)基因的重组腺病毒载体并对载体进行包装、收获病毒上清及纯化。将重组病毒液直接转染至 615小鼠皮下已接种MFC胃癌细胞成瘤的肿瘤部位进行抑瘤实验。结果 含CD基因的重组腺病毒载体经酶切鉴定正确 ;包装纯化后 ,检测病毒滴度为 1.2× 10 12 颗粒 /ml。将重组病毒液直接转染至肿瘤部位后 ,在前药 5 氟胞嘧啶 ( 5 FC)作用下 ,肿瘤生长较对照组明显缩小 (P <0 .0 1)。病理学检查可见肿瘤细胞大片坏死 ,且未转基因部位亦存在坏死区。结论 腺病毒载体转导的CD基因 ,不仅提高了转染效率 ,而且可以大幅度杀伤肿瘤细胞以及产生强效的旁观者效应。
Objective To improve the efficacy of gene delivering and to search for an effective way to kill the gastric cancer cells by suicidal gene. Method Firstly, the adenovirus combinatant vector containing cytosine deaminase (CD) gene was cloned and the combinatant plasmid was packed, amplified and purified. The MFC were injected subcutaneously into the flank of 615 mice to establish tumor models, and when tumors were palpable, 0.2ml adenovirus combinatant liquid containing CD gene was directly injected into xenografts following administration of 5 FC for 35 days. Results The positive clones selected by using endonuclease to digest the combinatants and the concentration of viral liquids containing CD gene was 1.2×10 12 particles/ml. Meaningfully, the antitumor effect was observed in tumors generated from MFC CD(+) tumors when the animals were given 5 FC by intraperitoneal injection. Pathologically, the tumors generated from MFC -CD(+) cells had hemorrhagic necrosis in large scale within the tumor regions as compared with that of untreated animals with few necrotic areas. Conclusion The improved anti tumor effect by adenovirus vector renders a basis for further research in gastric carcinoma involving the direct transduction of CD gene in tumors as well as induction of intensive bystander effect.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2000年第2期108-110,共3页
Chinese Journal of Digestion
基金
国家自然科学基金!3 96 0 0 14 2
