A Cox-2 promoter-based replication-selective adenoviral vector to target the Cox-2-expressing human bladder cancer cells
A Cox-2 promoter-based replication-selective adenoviral vector to target the Cox-2-expressing human bladder cancer cells
复制标题
DOI：
10.1158/1078-0432.ccr-03-0267
复制
发表时间：
2004-07-01
影响因子：
11.5
通讯作者：
Gotoh, A
中科院分区：
文献类型：
--
作者：
Shirakawa, T;Hamada, K;Gotoh, A
目的：环氧合酶-2（考克斯-2）是一种催化三尖杉酯碱合成的酶，在多种癌前病变和恶性病变（包括膀胱癌）中过表达。在本研究中，我们检查了使用基于考克斯-2启动子的复制选择性腺病毒靶向表达考克斯-2转录活性的膀胱癌细胞的可行性。一系列人类癌细胞系，包括三种膀胱癌细胞系（KK 47、T24和5637）的考克斯-2和CAR（柯萨奇病毒和腺病毒受体）mRNA表达水平。制备了E1 a表达受考克斯-2启动子调控的复制选择性腺病毒AdE 3-cox 2 -327，并在体外和体内检测了其组织特异性活性。三种膀胱癌细胞系比人前列腺癌细胞系PC 3、原代培养的人良性前列腺成纤维细胞PF细胞表达更高水平的考克斯-2 mRNA，和人结肠癌细胞系Colo 320。分别在KK 47、5637和Colo 320中检测到CAR mRNA的表达相对高于T24、PC-3和PF细胞。体外试验显示，与其他缺乏考克斯-2表达和/或CAR表达的细胞相比，表达考克斯-2和CAR的膀胱癌细胞KK 47和5637的生长受到显著抑制。结论：本研究证明了AdE 3-cox 2 -327（一种选择性复制的腺病毒）在体外和体内对表达考克斯-2的膀胱癌细胞的特异性和有效性。我们还发现，CAR在靶癌细胞中的表达是基于选择性复制的腺病毒的基因治疗有效性的重要因素。
Purpose: Cyclooxygenase-2 (Cox-2), an enzyme that catalyzes the synthesis of prostaglandins, is overexpressed in a variety of premalignant and malignant conditions, including urinary bladder cancer. In the present study, we examined the feasibility of using Cox-2 promoter-based replication-selective adenovirus for targeting bladder cancer cells that express Cox-2 transcriptional activity.Experimental Design: A series of human cancer cell lines, including three bladder cancer cell lines (KK47, T24, and 5637), were evaluated for their Cox-2 and CAR (the Coxsackievirus and adenovirus receptor) mRNA expression levels by quantitative real-time PCR. AdE3-cox2-327, a replication-selective adenovirus in which the expression of E1a is controlled by the Cox-2 promoter, was generated, and its tissue-specific activity was tested in vitro and in vivo.Results: Three bladder cancer cell lines express higher levels of Cox-2 mRNA than does the human prostate cancer cell line PC3, the primary cultured human benign prostatic fibroblast, PF cells, and the human colon cancer cell line Colo320. Relatively higher expression of CAR mRNA was detected in the KK47, 5637, respectively, and Colo320 than in the T24, PC-3, and PF cells. In vitro assays revealed significant growth suppression of both Cox-2- and CAR-expressing bladder cancer cells KK47 and 5637 in comparison with the other cells that lack Cox-2 expression and/or CAR expression.Conclusions: The present study demonstrated both specificity and efficacy of AdE3-cox2-327, a selectively replicated adenovirus, toward the Cox-2-expressing bladder cancer cells in vitro and in vivo. We also found that CAR expression in the target cancer cells is an important factor for the efficacy of selectively replicated adenovirus-based gene therapy.