ABSTRACT
BACKGROUND: Aldo-keto reductase (AKR) 1C family member 3 (AKR1C3), one of four identified human AKR1C enzymes, catalyzes steroid, prostaglandin, and xenobiotic metabolism. In the prostate, AKR1C3 is up-regulated in localized and advanced prostate adenocarcinoma, and is associated with prostate cancer (PCa) aggressiveness. Here we propose a novel pathological function of AKR1C3 in tumor angiogenesis and its potential role in promoting PCa progression. METHODS: To recapitulate elevated AKR1C3 expression in cancerous prostate, the human PCa PC-3 cell line was stably transfected with an AKR1C3 expression construct to establish PC3-AKR1C3 transfectants. Microarray and bioinformatics analysis were performed to identify AKR1C3-mediated pathways of activation and their potential biological consequences in PC-3 cells. Western blot analysis, reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and an in vitro Matrigel angiogenesis assays were applied to validate the pro-angiogenic activity of PC3-AKR1C3 transfectants identified by bioinformatics analysis. RESULTS: Microarray and bioinformatics analysis suggested that overexpression of AKR1C3 in PC-3 cells modulates estrogen and androgen metabolism, activates insulin-like growth factor (IGF)-1 and Akt signaling pathways, as well as promotes tumor angiogenesis and aggressiveness. Levels of IGF-1 receptor (IGF-1R) and Akt activation as well as vascular endothelial growth factor (VEGF) expression and secretion were significantly elevated in PC3-AKR1C3 transfectants in comparison to PC3-mock transfectants. PC3-AKR1C3 transfectants also promoted endothelial cell (EC) tube formation on Matrigel as compared to the AKR1C3-negative parental PC-3 cells and PC3-mock transfectants. Pre-treatment of PC3-AKR1C3 transfectants with a selective IGF-1R kinase inhibitor (AG1024) or a non-selective phosphoinositide 3-kinases (PI3K) inhibitor (LY294002) abolished ability of the cells to promote EC tube formation. CONCLUSIONS: Bioinformatics analysis followed by functional genomics demonstrated that AKR1C3 overexpression promotes angiogenesis and aggressiveness of PC-3 cells. These results also suggest that AKR1C3-mediated tumor angiogenesis is regulated by estrogen and androgen metabolism with subsequent IGF-1R and Akt activation followed by VEGF expression in PCa cells.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Endothelial Cells/enzymology , Hydroxyprostaglandin Dehydrogenases/metabolism , Neovascularization, Pathologic/enzymology , Prostatic Neoplasms/enzymology , 3-Hydroxysteroid Dehydrogenases/genetics , Blotting, Western , Cell Line, Tumor , Cell Survival , Computational Biology , Dihydrotestosterone/metabolism , Disease Progression , Endothelial Cells/drug effects , Endothelial Cells/pathology , Enzyme-Linked Immunosorbent Assay , Estradiol/metabolism , Gene Expression Profiling/methods , Gene Expression Regulation, Neoplastic , Gene Regulatory Networks , Humans , Hydroxyprostaglandin Dehydrogenases/genetics , Insulin-Like Growth Factor I/metabolism , Male , Neovascularization, Pathologic/pathology , Oligonucleotide Array Sequence Analysis , Promoter Regions, Genetic , Prostatic Neoplasms/blood supply , Prostatic Neoplasms/genetics , Prostatic Neoplasms/pathology , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Proto-Oncogene Proteins c-akt/metabolism , Receptor, IGF Type 1/antagonists & inhibitors , Receptor, IGF Type 1/metabolism , Receptors, Calcitriol/metabolism , Retinoid X Receptors/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Transfection , Tumor Suppressor Protein p53/metabolism , Up-Regulation , Vascular Endothelial Growth Factor A/metabolismABSTRACT
BACKGROUND: Originating from Africa, India, and the Middle East, frankincense oil has been important both socially and economically as an ingredient in incense and perfumes for thousands of years. Frankincense oil is prepared from aromatic hardened gum resins obtained by tapping Boswellia trees. One of the main components of frankincense oil is boswellic acid, a component known to have anti-neoplastic properties. The goal of this study was to evaluate frankincense oil for its anti-tumor activity and signaling pathways in bladder cancer cells. METHODS: Frankincense oil-induced cell viability was investigated in human bladder cancer J82 cells and immortalized normal bladder urothelial UROtsa cells. Temporal regulation of frankincense oil-activated gene expression in bladder cancer cells was identified by microarray and bioinformatics analysis. RESULTS: Within a range of concentration, frankincense oil suppressed cell viability in bladder transitional carcinoma J82 cells but not in UROtsa cells. Comprehensive gene expression analysis confirmed that frankincense oil activates genes that are responsible for cell cycle arrest, cell growth suppression, and apoptosis in J82 cells. However, frankincense oil-induced cell death in J82 cells did not result in DNA fragmentation, a hallmark of apoptosis. CONCLUSION: Frankincense oil appears to distinguish cancerous from normal bladder cells and suppress cancer cell viability. Microarray and bioinformatics analysis proposed multiple pathways that can be activated by frankincense oil to induce bladder cancer cell death. Frankincense oil might represent an alternative intravesical agent for bladder cancer treatment.
Subject(s)
Antineoplastic Agents, Phytogenic/therapeutic use , Boswellia , Oils, Volatile/therapeutic use , Plant Extracts/therapeutic use , Urinary Bladder Neoplasms/drug therapy , Urinary Bladder Neoplasms/genetics , Urothelium/drug effects , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line/drug effects , Cell Line, Tumor/drug effects , Cell Survival/drug effects , Gene Expression Profiling , Gene Expression Regulation , Humans , Oligonucleotide Array Sequence Analysis/methods , Resins, Plant , Transcription Factors , Urothelium/cytologyABSTRACT
Human aldo-keto reductase (AKR) 1C3 is a monomeric cytoplasmic multifunctional enzyme that reduces ketosteroids, ketoprostaglandins, and lipid aldehydes. AKR1C3 was initially identified as an enzyme involved in steroid metabolism. However, immunohistochemistry has demonstrated AKR1C3 in normal adult kidneys with expression in Bowman' capsule, the mesangial cells, proximal and distal tubules, as well as mature urothelial epithelium. The significance of its spatial distribution and metabolic activities in the kidney remains undefined. In addition to its ability to catalyze steroid hormones (including androgen, desoxycorticosterone, and progesterone) and involvement in prostaglandins metabolism, we suspect that AKR1C3 may function as a chemical barrier in the renal tubules for normal function in mature kidneys. Moreover, AKR1C3 may represent a developmental marker for some urological epithelial tissues. In this study, we demonstrate widespread expression of AKR1C3 in renal neoplasms with a phenotype recapitulating mature kidney (i.e., renal cell carcinoma) and urothelium also known as transitional epithelium (i.e., papillary urothelial carcinoma), but noted limited AKR1C3 expression in renal neoplasms with a phenotype recapitulating embryonic kidneys (i.e., Wilms' tumor). Our results suggest that AKR1C3 may represent a developmental marker that is related to renal epithelium maturity.