Laboratory of Cell and Gene Therapy, Institute for Advanced Medical Sciences, Hyogo College of Medicine, Nishinomiya, Japan.
Urology. 2013 Apr;81(4):920.e7-11. doi: 10.1016/j.urology.2012.12.023. Epub 2013 Feb 8.
To assess the oncolytic effect of fiber-substituted conditionally replicating adenovirus type 5 (Ad5) F35 vector on human bladder cancer cell lines such as 253J, 5637, KK-47, T24, TCCSUP, and UMUC-3 cells.
Ad5F35 and Ad5 conditionally replicating adenovirus vectors containing the E1 gene controlled by the human midkine promoter (Ad5F35/MKp-E1 and Ad5/MKp-E1, respectively) were constructed. Reverse transcriptase-polymerase chain reaction and cell viability assay were performed in cells transfected with Ad5F35/MKp-E1 or Ad5/MKp-E1.
Of the bladder cancer cells used, considerably lower expression of mRNA for Coxsackie and adenovirus receptor, an Ad5 receptor, was found with T24 and TCCSUP cells. However, the mRNA for CD46, an Ad35 receptor, was abundantly expressed in all the cell types. Ad5F35/MKp-E1 induced oncolysis in a plaque formation unit-dependent manner for all the bladder cancer cells used, with greater efficacy than Ad5/MKp-E1 for T24, TCCSUP, and 253J cells.
The results of the present study have shown that Ad5F35/MKp-E1 is more useful for the gene therapy of bladder cancer than Ad5/MKp-E1 is for some cell lines.
评估纤维替代条件复制腺病毒 5 型(Ad5)F35 载体对人膀胱癌细胞系(如 253J、5637、KK-47、T24、TCCSUP 和 UMUC-3 细胞)的溶瘤作用。
构建了携带人类中期因子启动子控制的 E1 基因的 Ad5F35 和条件复制腺病毒载体(分别为 Ad5F35/MKp-E1 和 Ad5/MKp-E1)。用 Ad5F35/MKp-E1 或 Ad5/MKp-E1 转染细胞后进行逆转录-聚合酶链反应和细胞活力测定。
在所使用的膀胱癌细胞中,T24 和 TCCSUP 细胞 Coxsackie 和腺病毒受体(Ad5 受体)的 mRNA 表达明显较低。然而,所有细胞类型均大量表达 CD46,即 Ad35 受体的 mRNA。Ad5F35/MKp-E1 以噬菌斑形成单位依赖性方式诱导所有使用的膀胱癌细胞发生溶瘤作用,对 T24、TCCSUP 和 253J 细胞的疗效优于 Ad5/MKp-E1。
本研究结果表明,与 Ad5/MKp-E1 相比,Ad5F35/MKp-E1 对一些细胞系的膀胱癌基因治疗更有用。