Lung tumorigenesis induced by human vascular endothelial growth factor (hVEGF)-A165 overexpression in transgenic mice and amelioration of tumor formation by miR-16.
Many studies have shown that vascular endothelial growth factor (VEGF), especially the human VEGF-A165 (hVEGF-A165) isoform, is a key proangiogenic factor that is overexpressed in lung cancer. We generated transgenic mice that overexpresses hVEGF-A165 in lung-specific Clara cells to investigate the development of pulmonary adenocarcinoma. In this study, three transgenic mouse strains were produced by pronuclear microinjection, and Southern blot analysis indicated similar patterns of the foreign gene within the genomes of the transgenic founder mice and their offspring. Accordingly, hVegf-A165 mRNA was expressed specifically in the lung tissue of the transgenic mice. Histopathological examination of the lung tissues of the transgenic mice showed that hVEGF-A165 overexpression induced bronchial inflammation, fibrosis, cysts, and adenoma. Pathological section and magnetic resonance imaging (MRI) analyses demonstrated a positive correlation between the development of pulmonary cancer and hVEGF expression levels, which were determined by immunohistochemistry, qRT-PCR, and western blot analyses. Gene expression profiling by cDNA microarray revealed a set of up-regulated genes (hvegf-A165, cyclin b1, cdc2, egfr, mmp9, nrp-1, and kdr) in VEGF tumors compared with wild-type lung tissues. In addition, overexpressing hVEGF-A165 in Clara cells increases CD105, fibrogenic genes (collagen α1, α-SMA, TGF-β1, and TIMP1), and inflammatory cytokines (IL-1, IL-6, and TNF-α) in the lungs of hVEGF-A165-overexpressing transgenic mice as compared to wild-type mice. We further demonstrated that the intranasal administration of microRNA-16 (miR-16) inhibited lung tumor growth by suppressing VEGF expression via the intrinsic and extrinsic apoptotic pathways. In conclusion, hVEGF-A165 transgenic mice exhibited complex alterations in gene expression and tumorigenesis and may be a relevant model for studying VEGF-targeted therapies in lung adenocarcinoma.
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.
Yu-Tang
YT
Tung
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.
Pin-Wu
PW
Huang
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.
Yu-Ching
YC
Chou
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.
Cheng-Wei
CW
Lai
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.
Hsiu-Po
HP
Wang
Department of Medicine, China Medical University Hospital, Taichung 404, Taiwan.
Heng-Chien
HC
Ho
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.|Department of Internal Medicine, China Medical University Hospital, Taichung 404, Taiwan.
Chih-Ching
CC
Yen
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.|Department of Internal Medicine, China Medical University Hospital, Taichung 404, Taiwan.
Chih-Yen
CY
Tu
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.
Tung-Chou
TC
Tsai
Department of General Surgery and Department of Internal Medicine, Taichung Veterans General Hospital, Taichung 407, Taiwan.
Dah-Cherng
DC
Yeh
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.|Division of Chest Medicine, Department of Internal Medicine, Taichung Veterans General Hospital, Taichung 407, Taiwan.
Jiun-Long
JL
Wang
Department of Medical Biotechnology and Laboratory Sciences, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.|Molecular Medicine Research Center, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan.
Kowit-Yu
KY
Chong
Department of Life Sciences and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.|Rong-Hsing Translational Medicine Center and iEGG Center, National Chung Hsing University, Taichung 402, Taiwan.
Chuan-Mu
CM
Chen
D000818:Animals; D063646:Carcinogenesis; D045744:Cell Line, Tumor; D002642:Chick Embryo; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D008279:Magnetic Resonance Imaging / Q000379:methods; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000008:administration & dosage / Q000235:genetics*; D015514:Oncogene Proteins, Fusion / Q000096:biosynthesis / Q000235:genetics; D015398:Signal Transduction; D059467:Transcriptome; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
C505362:MIRN16 microRNA, human; D035683:MicroRNAs; D015514:Oncogene Proteins, Fusion; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
VEGF; magnetic resonance imaging (MRI); miRNA therapy; pulmonary tumorigenesis; transgenic mice