Beneficial Oncolytic Effect of Fiber-substituted Conditionally Replicating Adenovirus on Human Lung Cancer

• Published in: Anticancer research 2012-11, Vol.32 (11), p.4891-4895
• Main Authors: KANNO, Takeshi, GOTOH, Akinobu, NAKANO, Takashi, TAGAWA, Masatoshi, NISHIZAKI, Tomoyuki
• Format: Article
• Language: English
• Subjects: Adenocarcinoma; Adenoviridae - genetics; Adenovirus; Biological and medical sciences; Blotting, Western; CD46 antigen; Cell Line, Tumor; Cell surface; Cytokines - administration & dosage; Cytokines - genetics; E1 gene; Early region; Embryos; Expression vectors; Gene expression; Gene therapy; Genetic Therapy - methods; Genetic Vectors; Humans; Infection; Lung cancer; Lung Neoplasms; Medical sciences; midkine; Oncolysis; Oncolytic Virotherapy - methods; Pneumology; Polymerase chain reaction; Promoters; Reverse Transcriptase Polymerase Chain Reaction; Transfection; Tumor cell lines; Tumors; Tumors of the respiratory system and mediastinum; Western blotting
• ISSN: 0250-7005; 1791-7530

Adenovirus vectors have been utilized for a variety of cancer gene therapy. The present study examined the oncolytic effect of adenovirus type 5 (Ad5) and fiber-substituted conditionally replicating adenovirus (CRAD) Ad5/F35 vectors on human lung cancer A549 cells (an epithelial adenocarcinoma cell line), SBC-3 cells (a small-cell cancer cell line), and Lu-65 cells (a giant-cell lung cancer cell line). For adenovirus, the first mRNA/protein to be made (~1 h after infection) is early region 1A (E1A). Ad5F35 and Ad5 CRAD vectors containing the E1 gene, controlled by the human midkine promoter (Ad5F35/MKp-E1 and Ad5/MKp-E1, respectively) were constructed. Reverse transcription-polymerase chain reaction (RT-PCR), western blotting and cell viability assays were carried out in cells transfected with Ad5/MKp-E1 and Ad5F35/MKp-E1. Less expression of mRNA and protein for coxsackie and adenovirus receptor (CAR), a cell surface target of Ad5, was found with lung cancer cells as compared with the expression in human embryonic kidney 293 (HEK293) cells, but otherwise mRNA and CD46 protein, a cell surface target of Ad35, was expressed in lung cancer cells as much as in HEK293 cells. Both Ad5/MKp-E1 and Ad5F35/MKp-E1 induced oncolysis of lung cancer cells in a viral particle-dependent manner, with more efficient advantage for Ad5F35/MKp-E1. The results of the present study suggest that Ad5F35/MKp-E1 is more useful for the gene therapy of human lung cancer than Ad5/MKp-E1 is.