携带Kringle 5基因重组腺病毒的构建及其对荷肺癌小鼠生存期的作用  

Construction of recombinant adenovirus carrying Kringle 5 gene and effects on the survival of pulmonary carcinoma bearing mice

作　　者：刘岩[1] 刘芳[2] 叶迅[2] 董继斌[3] 陆琴[2] 赵毅[2] 李秋华[1] 陈红专[2] 

机构地区：[1]复旦大学附属华山医院眼科,上海200040 [2]上海交通大学医学院药理学教研室,上海200025 [3]复旦大学药学院生物化学教研室,上海200032

出　　处：《中国临床药理学与治疗学》2008年第3期254-258,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：上海市自然科学基金项目(03Z214116)

摘　　要：目的:构建携带人纤溶酶原Kringle 5(K5)基因重组腺病毒并观察其在小鼠肺癌细胞中的表达和对荷TC-1瘤小鼠的生存期的影响。方法:构建携带人纤溶酶原K5基因表达框的腺病毒载体Ad-EF1α-K5,用RT-PCR法检测腺病毒感染TC-1细胞后K5 mRNA的表达,用免疫沉淀和Western bloting法检测腺病毒感染TC-1细胞后K5蛋白的表达。建立荷TC-1小鼠肺癌C57BL/6小鼠模型,观察重组腺病毒对荷瘤小鼠生存期的影响。取肿瘤组织做免疫组织化学染色并计数微血管密度。结果:携带K5基因的人V型腺病毒能有效感染小鼠肺癌TC-1细胞,在转录和翻译水平进行表达,能有效延长荷瘤鼠的生存期并降低肿瘤微血管密度。结论:腺病毒可作为便捷的载体携带K5基因感染肿瘤细胞,分泌表达K5蛋白,延长荷瘤小鼠生存期,可能与其通过抑制肿瘤区血管生成、遏制肿瘤区血供有关。AIM： To construct the recombinant adenovirus containing human plasminogen Kringle 5 （K5） gene, then observe the secreted expression and effect of K5 protein on TC-1 tumors. METHODS： We cloned K5 gene into the genome of replication-defective adenovirus by virus recombination technology; the K5 RNA expression was evaluated by RT-PCR; the K5 protein secreted expression was determined by Immunoprecipitation and Western blotting; the effect of K5 was observed in xenograft lung cancr mice model; and the tumor microvascular density （MVD） was counted. RESULTS： The recombined adenovirus Ad-EF1α-K5 was constructed successfully and was expressing in the levels of both mRNA and protein. In the mice lung cancer xenograft model, the recombinant adenovirus can delay the growth of established TC-1 tumors and decrease the MVD. CONCLUSION： Recombinant adenovirus vectors Ad-EF1α-K5 can efficiently express protein K5 and inhibit tumor growth in the mice lung cancer xenograft model. The effect may related to inhibitory effect on angiogenesis.

关 键 词：腺病毒 KRINGLE 5 血管生成 TC-1 

分 类 号：R979.19[医药卫生—药品]