Conclusion Taking together, we have generated a novel oncolytic adenoviral vector in which the main difference with currently used oncolytic
adenoviral FHPI vector ONYX-015 is hTERT controlled replication and armed with HSV-TK. The hTERT promoter used in this study is high stringency and provide the base for tumor-specific replication. Ad.hTERT-E1A-TK itself was able to inhibit tumor growth thanks to its replicative ability and oncolytic effect. Moreover, its tumor killing effect could be further enhanced by prodrug GCV. Our study showed that Ad.hTERT-E1A-TK/GCV could efficiently kill NSCLC tumor cells both in vitro and in vivo. Therefore, we concluded that Ad.hTERT-E1A-TK, as a potent and safe antitumor strategy, could provide a potential new option for NSCLC biotherapy. Acknowledgements This work was Mocetinostat in vitro supported by Grant AZD5363 manufacturer of National Basic Research Project of China (2010CB529902), National High-tech R&D program (2007AA021202), National Natural Science Foundation for Outstanding Youth (30325043). Electronic supplementary material Additional file 1: Schematic diagram of Ad.hTERT-E1A-CD or Ad.hTERT-E1A-TK adenoviral construct. Ad.hTERT-E1A-CD or Ad.hTERT-E1A-TK adenoviral vector had been constructed in the way described in this figure. ITR, inverted repeats of the adenovirus genome; ΔE1 and ΔE3, E1 and E3 region deleted. (TIFF 73 KB) Additional file 2: Western blotting analysis of TK gene expression. NCIH460
Cells were infected with Ad-hTERT-E1A-TK at a MOI of 10. Cell lysates were harvested 48 h later, and
immunobloted by anti HA-tag antibody. NCIH460 Cells which had been transfected Sclareol with plasmid containing TK gene were used as positive control, and uninfected NCIH460 cells were used as negative control. (TIFF 667 KB) Additional file 3: Tumor cell killing effect of Ad.hTERT-E1A-TK on different tumor cells. Crystal violet staining of tumor cells after infection with different adenoviral vectors. SW1990, SMMC-7721 and HeLa cells were plated into 24-well plates and treated with different dose of adenoviral vectors or prodrug or untreated as indicated in figure. 5 days later the plates were stained with crystal violet. (TIFF 2 MB) References 1. Lee CB, Stinchcombe TE, Rosenman JG, Socinski MA: Therapeutic advances in local-regional therapy for stage III non-small-cell lung cancer: evolving role of dose-escalated conformal (3-dimensional) radiation therapy. Clin Lung Cancer 2006, 8:195–202.PubMedCrossRef 2. Rossi A, Maione P, Colantuoni G, Ferrara C, Rossi E, Guerriero C, Nicolella D, Falanga M, Palazzolo G, Gridelli C: Recent developments of targeted therapies in the treatment of non-small cell lung cancer. Curr Drug Discov Technol 2009, 6:91–102.PubMedCrossRef 3. Ricciardi S, Tomao S, Marinis F: Toxicity of targeted therapy in non-small-cell lung cancer management. Clin Lung Cancer 2009, 10:28–35.PubMedCrossRef 4. Herbst RS, Sandler AB: Overview of the current status of human epidermal growth factor receptor inhibitors in lung cancer.