【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。更多还原

【Abstract】 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.更多还原