Cancer dual gene therapywith oncolytic adenoviruses expressing TRAIL and IL-12 transgenes markedly eradicatedhuman hepatocellular carcinoma both in vitro and in vivo
Adel GalalAhmed El-Shemi*, Ahmad Ashshi, Mohammed Basalamah,Youjin Na, and Chae-Ok YUN
*Corresponding Author:
Dr. Adel Galal El-Shemi, Department of Laboratory Medicine Department, Faculty of Applied Medical Sciences, Umm Al-Qura University, Makkah, PO Box 7607, Saudi Arabia. Tel: +966 509655135; Fax: +966 12 5270000 Ext: 4242; email:
Funding Source: This project was funded by the National Science, Technology and Innovation Plan (MARRIFAH) - King Abdul Aziz City for Science and Technology (KACST), the Kingdom of Saudi Arabia, Award Number (11-MED2065-10).
ABSTRACT
BACKGROUND: Cancer gene therapy-mediated by oncolytic adenoviruses (Ads) encodingimmunostimulatoryinterleukin-12 (IL-12) gene, or pro-apoptotic tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene, have been recently emerged as a novel strategy in cancer therapy. In this study, we generated, and the investigated the single and combination therapy of two oncolytic Ad encoding human TRAIL gene (Ad-ΔB/TRAIL) and oncolytic Ad encoding human IL-12 gene (Ad-ΔB/IL-12) on human hepatocellular carcinoma ( human HCC) cell lines and on orthotopic human HCC (Hep3B) model in athymicnude mice.
RESULTS: Ad-ΔB/TRAIL and Ad-ΔB/IL-12 combination therapy elicited a more significant cytopathic effect on HCC cellsand additive growth suppression of the xenogarfted tumor compared with their single therapy, without overlapping toxicity. Additionally, the augmented anti-HCC activity of Ad-ΔB/TRAIL and Ad-ΔB/IL-12 combination therapy had also resulted in a more activation of the apoptotic-caspase-3 and-8 pathway, overproduction ofinterferon gamma (IFN-γ), high infiltration rate of natural killer cells and antigen presenting cells. Moreover, Ad-ΔB/TRAIL and Ad-ΔB/IL-12 combination treatment additively reduced vascular endothelial growth factor (VEGF) and CD31 expression as well as the microvessel density in the tumor tissues.
Conclusions:Cancer dual gene therapy with Ad-ΔB/TRAIL and Ad-ΔB/IL-12 was synergistically interacted in suppressing human HCC in vitro and in vivo with enhanced activation of anti-tumor immunity and apoptosis, and also enhanced inhibition of tumor angiogenesis and vasculature.
Figure 1. Representative photographs of histological, immunohistochemical and TUNEL analyses of the tumor tissues. H&E: hematoxylin and eosin staining for histopathology; TUNEL: staining of apoptotic cells in tumor tissue; NK1.1:to detect the infiltrated natural killer (NK) cells,the tumor tissues were treated with purified anti-mouse NK1.1 monoclonal antibody; CD11b: to detect the recruited antigen-presenting cells (dendritic cells and macrophage), the tumor tissues were treated with purified anti-mouse CD11b monoclonal antibody; and CD31: to detect tumor microvessels endothelial cells and vascular density, the tumor tissues were treated with anti-CD31 antibody. Brown staining indicates positive staining cells.