title: Identification of pro-inflammatory cytokines associated with muscle invasive bladder cancer; the roles of IL-5, IL-20, and IL-28A.
abstract: We used gene expression profiling to identify inflammatory cytokines that correlate with bladder cancer development. Gene expression profiles of the tissue samples were investigated using cDNA microarrays that contained 103 non-muscle invasive bladder cancers (NMIBC), 62 muscle invasive bladder cancers (MIBC), 58 samples of histologically normal-looking surrounding tissues, and 10 normal, healthy subjects who served as the control cohort for comparison. We grouped the data-sets according to biological characterizations and focused on immune response genes with at least 2-fold differential expression in MIBC vs. controls. The experimental data-set identified 36 immune-related genes that were significantly altered in MIBC samples. In addition, 10 genes were up-regulated and 26 genes were down-regulated in MIBC samples compared with the normal tissues. Among the 10 up-regulated molecules examined, the capacity for both wound-healing migration and invasion was enhanced in response to IL-5, IL-20, and IL-28A in bladder cancer cell lines (253J and EJ cells), compared with untreated cells. The expression levels of IL-5, IL-20, and IL-28A were increased in patients with MIBC. All 3 cytokines and their receptors were produced in bladder cancer cell lines, as determined by real-time PCR, immunoblot analysis and confocal immunofluorescence. Up-regulation of MMP-2 and MMP-9 was found after IL-5, IL-20, and IL-28A stimulation in both cell types. Moreover, an EMSA assay showed that treatment with IL-5, IL-20, and IL-28A induced activation of the transcription factors NF-κB and AP-1 that regulate the MMP-9 promoter. Finally, activation of MAPK and Jak-Stat signaling was observed after the addition of IL-5, IL-20, and IL-28A to bladder cancer cells. This study suggests the presence of specific inflammatory cytokine (IL-5, IL-20, and IL-28A)-mediated association in bladder cancer development. All 3 cytokines may be important new molecular targets for the modulation of migration and invasion in bladder cancer.
authors.affiliation: Department of Biotechnology, Chungju National University, Chungju, Chungbuk, South Korea.
authors.forename: Se-Jung
authors.initials: SJ
authors.lastname: Lee
authors.forename: Eo-Jin
authors.initials: EJ
authors.lastname: Lee
authors.forename: Seon-Kyu
authors.initials: SK
authors.lastname: Kim
authors.forename: Pildu
authors.initials: P
authors.lastname: Jeong
authors.forename: Young-Hwa
authors.initials: YH
authors.lastname: Cho
authors.forename: Seok Joong
authors.initials: SJ
authors.lastname: Yun
authors.forename: Sangtae
authors.initials: S
authors.lastname: Kim
authors.forename: Gi-Young
authors.initials: GY
authors.lastname: Kim
authors.forename: Yung Hyun
authors.initials: YH
authors.lastname: Choi
authors.forename: Eun-Jong
authors.initials: EJ
authors.lastname: Cha
authors.forename: Wun-Jae
authors.initials: WJ
authors.lastname: Kim
authors.forename: Sung-Kwon
authors.initials: SK
authors.lastname: Moon
mesh_terms: D000368:Aged; D045744:Cell Line, Tumor; D002465:Cell Movement; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000276:immunology; D006801:Humans; D015848:Interleukin-5 / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology; D007378:Interleukins / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D009130:Muscle, Smooth / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D009361:Neoplasm Invasiveness; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D000095022:Interferon Lambda
chemical_list: D015848:Interleukin-5; D007378:Interleukins; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; C000624353:interferon-lambda, human; D000095022:Interferon Lambda; C508599:IL5 protein, human; C422042:interleukin 20