Inhibitory effect of Survivin promoter-regulated oncolytic adenovirus carrying P53 gene against gallbladder cancer
Inhibitory effect of Survivin promoter-regulated oncolytic adenovirus carrying P53 gene against gallbladder cancer
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Survivin启动子调控的P53基因溶瘤腺病毒对胆囊癌的抑制作用
DOI：
10.1016/j.molonc.2011.10.001
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发表时间：
2011-12-01
影响因子：
6.6
通讯作者：
Jiang, Xiaoqing
中科院分区：
文献类型：
--
作者：
Liu, Chen;Sun, Bin;Jiang, Xiaoqing
基因治疗已成为治疗恶性肿瘤的重要策略，但基因转移效率低、转基因表达差、靶向性有限等问题严重制约了肿瘤基因治疗的临床应用。胆囊癌具有进展快、预后差、Survivin异常高表达等特点。本研究利用肿瘤特异性Survivin启动子调控的携带P53基因的溶瘤腺病毒载体，构建了广谱、特异、安全、有效的基因-病毒治疗系统AdSurp-P53。通过检测增强型绿色荧光蛋白（EGFP）、E1 A和靶基因P53在溶瘤腺病毒系统中的表达，证实Survivin启动子调控的溶瘤腺病毒在Survivin阳性胆囊癌细胞中具有高增殖活性和高P53表达。体外细胞毒实验表明，AdSurp-P53对胆囊癌细胞和肝癌细胞具有较强的细胞毒作用。感染复数为1 pfu/cell的AdSurp-P53感染EH-GB 1细胞后，细胞存活率低于40%，而相同感染复数的Ad-P53感染EH-GB 1细胞后，细胞存活率高于90%，表明AdSurp-P53对EH-GB 1细胞具有较强的细胞毒作用。当AdSurp-P53的总剂量为1 × 10（9）pfu时，EH-GB 1裸鼠移植瘤的生长受到明显抑制，末端dUTP缺口末端标记法（TUNEL）显示癌细胞的凋亡率为（33.4 ± 8.4）%。这种溶瘤腺病毒系统克服了基因治疗长期存在的缺陷：转基因表达差，只能靶向特定的肿瘤，其治疗效果优于已上市的传统Ad-P53治疗方案;我们的系统可能用于晚期胆囊癌和其他癌症，对化疗，放疗不敏感或失去手术治疗机会的患者。(C)2011年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Gene therapy has become an important strategy for treatment of malignancies, but problems remains concerning the low gene transferring efficiency, poor transgene expression and limited targeting specific tumors, which have greatly hampered the clinical application of tumor gene therapy. Gallbladder cancer is characterized by rapid progress, poor prognosis, and aberrantly high expression of Survivin. In the present study, we used a human tumor-specific Survivin promoter-regulated oncolytic adenovirus vector carrying P53 gene, whose anti-cancer effect has been widely confirmed, to construct a wide spectrum, specific, safe, effective gene-viral therapy system, AdSurp-P53. Examining expression of enhanced green fluorecent protein (EGFP), E1A and the target gene P53 in the oncolytic adenovirus system validated that Survivin promoter-regulated oncolytic adenovirus had high proliferation activity and high P53 expression in Survivin-positive gallbladder cancer cells. Our in vitro cytotoxicity experiment demonstrated that AdSurp-P53 possessed a stronger cytotoxic effect against gallbladder cancer cells and hepatic cancer cells. The survival rate of EH-GB1 cells was lower than 40% after infection of AdSurp-P53 at multiplicity of infection (MOI) = 1 pfu/cell, while the rate was higher than 90% after infection of Ad-P53 at the same MOI, demonstrating that AdSurp-P53 has a potent cytotoxicity against EH-GB1 cells. The tumor growth was greatly inhibited in nude mice bearing EH-GB1 xenografts when the total dose of AdSurp-P53 was 1 x 10(9) pfu, and terminal dUTP nick end-labeling (TUNEL) revealed that the apoptotic rate of cancer cells was (33.4 +/- 8.4)%. This oncolytic adenovirus system overcomes the long-standing shortcomings of gene therapy: poor transgene expression and targeting of only specific tumors, with its therapeutic effect better than the traditional Ad-P53 therapy regimen already on market; our system might be used for patients with advanced gallbladder cancer and other cancers, who are not sensitive to chemotherapy, radiotherapy, or who lost their chance for surgical treatment. (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.