miR-143 and miR-145 are downregulated in ulcerative colitis: Putative regulators of inflammation and protooncogenes*
Corresponding Author
Joel R. Pekow MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
900 East 57th St, MB#9, Chicago, IL 60637Search for more papers by this authorUrszula Dougherty MS
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorReba Mustafi PhD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorHongyan Zhu MD, PhD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorMasha Kocherginsky PhD
Department of Health Studies, University of Chicago, Chicago, Illinois
Search for more papers by this authorDavid T. Rubin MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorStephen B. Hanauer MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorJohn Hart MD
Department of Pathology, University of Chicago, Chicago, Illinois
Search for more papers by this authorEugene B. Chang MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorAlessandro Fichera MD
Department of Surgery, University of Chicago, Chicago, Illinois
Search for more papers by this authorLoren J. Joseph MD
Department of Pathology, University of Chicago, Chicago, Illinois
Search for more papers by this authorMarc Bissonnette MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorCorresponding Author
Joel R. Pekow MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
900 East 57th St, MB#9, Chicago, IL 60637Search for more papers by this authorUrszula Dougherty MS
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorReba Mustafi PhD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorHongyan Zhu MD, PhD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorMasha Kocherginsky PhD
Department of Health Studies, University of Chicago, Chicago, Illinois
Search for more papers by this authorDavid T. Rubin MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorStephen B. Hanauer MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorJohn Hart MD
Department of Pathology, University of Chicago, Chicago, Illinois
Search for more papers by this authorEugene B. Chang MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorAlessandro Fichera MD
Department of Surgery, University of Chicago, Chicago, Illinois
Search for more papers by this authorLoren J. Joseph MD
Department of Pathology, University of Chicago, Chicago, Illinois
Search for more papers by this authorMarc Bissonnette MD
Section of Gastroenterology, Hepatology, and Nutrition, University of Chicago, Chicago, Illinois
Search for more papers by this authorSupported by the University of Chicago, Institute of Translational Medicine (to J.P.), Crohn's and Colitis Foundation of America (to J.P.), Crohn's Disease Working Group (to J.P.), Gastrointestinal Research Foundation of Chicago (to J.P.), University of Chicago, Comprehensive Cancer Center (to M.B.).
Abstract
Background:
miR-143 and miR-145 are believed to function as colon cancer tumor suppressors, as they inhibit colon cancer cell growth and are downregulated in sporadic colonic tumors. We speculated that miR-143 and miR-145 might also be downregulated and contribute to malignant transformation of colonic epithelium in longstanding ulcerative colitis (UC).
Methods:
Biopsies were obtained 20 cm proximal to the anus from individuals with quiescent UC and from normal controls. RNA and proteins were extracted and measured. miR-143 and miR-145 were quantified by real-time polymerase chain reaction (PCR) and miR-145 was also assessed by in situ hybridization. Putative targets of these miRNAs, K-RAS, API5, MEK-2 (miR-143), and IRS-1 (miR-145) were determined by western blotting. To assess the effects of miR-143 and miR-145 on these predicted targets, HCT116 and HCA-7 colorectal cancer cells were transfected with miR-143 and miR-145 and expression levels of these proteins were measured.
Results:
In UC, miR-143 and miR-145 were significantly downregulated 8.3-fold (3.4–20.1) (P < 0.0001) and 4.3-fold (2.3–7.8) (P < 0.0001), respectively, compared to normal colon. In contrast, IRS-1, K-RAS, API5, and MEK-2 were upregulated in UC, consistent with their assignments as targets of these miRNAs. Furthermore, transfected miR-143 and miR-145 significantly downregulated these proteins in HCT116 or HCA-7 cells.
Conclusions:
Compared to normal colonic mucosa, in chronic UC miR-143 and miR-145 were significantly downregulated and their predicted targets, IRS-1, K-RAS, API5, and MEK-2 were upregulated. We postulate that loss of these tumor suppressor miRNAs predispose to chronic inflammation and neoplastic progression in IBD. (Inflamm Bowel Dis 2011;)
REFERENCES
- 1 Eaden JA, Abrams KR, Mayberry JF. The risk of colorectal cancer in ulcerative colitis: a meta-analysis. Gut. 2001; 48: 526–535.
- 2 Rutter MD, Saunders BP, Wilkinson KH, et al. Thirty-year analysis of a colonoscopic surveillance program for neoplasia in ulcerative colitis. Gastroenterology. 2006; 130: 1030–1038.
- 3 Rutter M, Saunders B, Wilkinson K, et al. Severity of inflammation is a risk factor for colorectal neoplasia in ulcerative colitis. Gastroenterology. 2004; 126: 451–459.
- 4 Gupta RB, Harpaz N, Itzkowitz S, et al. Histologic inflammation is a risk factor for progression to colorectal neoplasia in ulcerative colitis: a cohort study. Gastroenterology. 2007; 133: 1099–1105; quiz 1340–1091.
- 5 Rubin DT. The changing face of colorectal cancer in inflammatory bowel disease: progress at last! Gastroenterology. 2006; 130: 1350–1352.
- 6 Itzkowitz SH. Molecular biology of dysplasia and cancer in inflammatory bowel disease. Gastroenterol Clin North Am. 2006; 35: 553–571.
- 7 van der Woude CJ, Kleibeuker JH, Jansen PL, et al. Chronic inflammation, apoptosis and (pre-)malignant lesions in the gastro-intestinal tract. Apoptosis. 2004; 9: 123–130.
- 8 Xie J, Itzkowitz SH. Cancer in inflammatory bowel disease. World J Gastroenterol. 2008; 14: 378–389.
- 9 Willenbucher RF, Aust DE, Chang CG, et al. Genomic instability is an early event during the progression pathway of ulcerative-colitis-related neoplasia. Am J Pathol. 1999; 154: 1825–1830.
- 10 Burmer GC, Rabinovitch PS, Haggitt RC, et al. Neoplastic progression in ulcerative colitis: histology, DNA content, and loss of a p53 allele. Gastroenterology. 1992; 103: 1602–1610.
- 11 Svec J, Musilkova J, Bryndova J, et al. Enhanced expression of proproliferative and antiapoptotic genes in ulcerative colitis-associated neoplasia. Inflamm Bowel Dis. 2010; 16: 1127–1137.
- 12 Lawrance IC, Fiocchi C, Chakravarti S. Ulcerative colitis and Crohn's disease: distinctive gene expression profiles and novel susceptibility candidate genes. Hum Mol Genet. 2001; 10: 445–456.
- 13 Noble CL, Abbas AR, Cornelius J, et al. Regional variation in gene expression in the healthy colon is dysregulated in ulcerative colitis. Gut. 2008; 57: 1398–1405.
- 14 Okahara S, Arimura Y, Yabana T, et al. Inflammatory gene signature in ulcerative colitis with cDNA macroarray analysis. Aliment Pharmacol Ther. 2005; 21: 1091–1097.
- 15 Wu F, Dassopoulos T, Cope L, et al. Genome-wide gene expression differences in Crohn's disease and ulcerative colitis from endoscopic pinch biopsies: insights into distinctive pathogenesis. Inflamm Bowel Dis. 2007; 13: 807–821.
- 16 Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004; 116: 281–297.
- 17 Liu W, Mao SY, Zhu WY. Impact of tiny miRNAs on cancers. World J Gastroenterol. 2007; 13: 497–502.
- 18 Ambros V. The functions of animal microRNAs. Nature. 2004; 431: 350–355.
- 19 O'Connell RM, Taganov KD, Boldin MP, et al. MicroRNA-155 is induced during the macrophage inflammatory response. Proc Natl Acad Sci U S A. 2007; 104: 1604–1609.
- 20 Volinia S, Calin GA, Liu CG, et al. A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci U S A. 2006; 103: 2257–2261.
- 21 Wu F, Zikusoka M, Trindade A, et al. MicroRNAs are differentially expressed in ulcerative colitis and alter expression of macrophage inflammatory peptide-2 alpha. Gastroenterology. 2008; 135: 1624–1635 e1624.
- 22 Chen X, Guo X, Zhang H, et al. Role of miR-143 targeting KRAS in colorectal tumorigenesis. Oncogene. 2009; 28: 1385–1392.
- 23 Akao Y, Nakagawa Y, Naoe T. MicroRNAs 143 and 145 are possible common onco-microRNAs in human cancers. Oncol Rep. 2006; 16: 845–850.
- 24 Starczynowski DT, Kuchenbauer F, Argiropoulos B, et al. Identification of miR-145 and miR-146a as mediators of the 5q-syndrome phenotype. Nat Med. 2010; 16: 49–58.
- 25 Fasseu M, Treton X, Guichard C, et al. Identification of restricted subsets of mature microRNA abnormally expressed in inactive colonic mucosa of patients with inflammatory bowel disease. PLoS One. 2010; 5: e13160.
- 26 Yang Y, Chaerkady R, Kandasamy K, et al. Identifying targets of miR-143 using a SILAC-based proteomic approach. Mol Biosyst. 2010; 6: 1873–1882.
- 27 Wang CJ, Zhou ZG, Wang L, et al. Clinicopathological significance of microRNA-31, -143 and -145 expression in colorectal cancer. Dis Markers. 2009; 26: 27–34.
- 28 Shi B, Sepp-Lorenzino L, Prisco M, et al. Micro RNA 145 targets the insulin receptor substrate-1 and inhibits the growth of colon cancer cells. J Biol Chem. 2007; 282: 32582–32590.
- 29
Zhu H,
Dougherty U,
Mustafi R, et al.
Putative tumor supressors miR-143 and miR-145 inhibit HCT1116 colon cancer cell growth in tumor xenografts: roles of K-RAS, MYC, Ccnd2, and CDK6.
Gastroenterology.
2010;
138:
S–93.
10.1016/S0016-5085(10)60424-6 Google Scholar
- 30 Shinozaki M, Watanabe T, Kubota Y, et al. High proliferative activity is associated with dysplasia in ulcerative colitis. Dis Colon Rectum. 2000; 43: S34–39.
- 31 Andersen SN, Rognum TO, Bakka A, et al. Ki-67: a useful marker for the evaluation of dysplasia in ulcerative colitis. Mol Pathol. 1998; 51: 327–332.
- 32 Arai N, Mitomi H, Ohtani Y, et al. Enhanced epithelial cell turnover associated with p53 accumulation and high p21WAF1/CIP1 expression in ulcerative colitis. Mod Pathol. 1999; 12: 604–611.
- 33 Krishna M, Woda B, Savas L, et al. Expression of p53 antigen in inflamed and regenerated mucosa in ulcerative colitis and Crohn's disease. Mod Pathol. 1995; 8: 654–657.
- 34 Silahtaroglu AN, Nolting D, Dyrskjot L, et al. Detection of microRNAs in frozen tissue sections by fluorescence in situ hybridization using locked nucleic acid probes and tyramide signal amplification. Nat Protoc. 2007; 2: 2520–2528.
- 35 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001; 25: 402–408.
- 36 Yuan JS, Reed A, Chen F, et al. Statistical analysis of real-time PCR data. BMC Bioinformatics. 2006; 7: 85.
- 37 Inoue T, Murano M, Kuramoto T, et al. Increased proliferation of middle to distal colonic cells during colorectal carcinogenesis in experimental murine ulcerative colitis. Oncol Rep. 2007; 18: 1457–1462.
- 38 Sebolt-Leopold JS, Dudley DT, Herrera R, et al. Blockade of the MAP kinase pathway suppresses growth of colon tumors in vivo. Nat Med. 1999; 5: 810–816.
- 39 Morris EJ, Michaud WA, Ji JY, et al. Functional identification of Api5 as a suppressor of E2F-dependent apoptosis in vivo. PLoS Genet. 2006; 2: e196.
- 40 Gunter MJ, Hayes RB, Chatterjee N, et al. Insulin resistance-related genes and advanced left-sided colorectal adenoma. Cancer Epidemiol Biomarkers Prev. 2007; 16: 703–708.
- 41
Pekow J,
Dougherty U,
Zhu H, et al.
MiR-143 and miR-145 are Differentially Expressed in the Right and Left Colon and Likely Regulate Differential Expression of K-RAS and Irs-1: Implications for Colon Cancer.
Gastroenterology.
2010;
138:
S-256.
10.1016/S0016-5085(10)61168-7 Google Scholar
- 42 Xie H, Lim B, Lodish HF. MicroRNAs induced during adipogenesis that accelerate fat cell development are downregulated in obesity. Diabetes. 2009; 58: 1050–1057.
- 43 Akao Y, Nakagawa Y, Hirata I, et al. Role of anti-oncomirs miR-143 and -145 in human colorectal tumors. Cancer Gene Ther. 2010; 17: 398–408.
- 44 La Rocca G, Badin M, Shi B, et al. Mechanism of growth inhibition by MicroRNA 145: the role of the IGF-I receptor signaling pathway. J Cell Physiol. 2009; 220: 485–491.
- 45 Bergmann U, Funatomi H, Kornmann M, et al. Increased expression of insulin receptor substrate-1 in human pancreatic cancer. Biochem Biophys Res Commun. 1996; 220: 886–890.
- 46 Ramocki NM, Wilkins HR, Magness ST, et al. Insulin receptor substrate-1 deficiency promotes apoptosis in the putative intestinal crypt stem cell region, limits Apcmin/+ tumors, and regulates Sox9. Endocrinology. 2008; 149: 261–267.
- 47 Chang Q, Li Y, White MF, et al. Constitutive activation of insulin receptor substrate 1 is a frequent event in human tumors: therapeutic implications. Cancer Res. 2002; 62: 6035–6038.
- 48 Jaiswal BS, Janakiraman V, Kljavin NM, et al. Combined targeting of BRAF and CRAF or BRAF and PI3K effector pathways is required for efficacy in NRAS mutant tumors. PLoS One. 2009; 4: e5717.
- 49 Bramis J, Zacharatos P, Papaconstantinou I, et al. E2F-1 transcription factor immunoexpression is inversely associated with tumor growth in colon adenocarcinomas. Anticancer Res. 2004; 24: 3041–3047.
- 50 Su DM, Zhang Q, Wang X, et al. Two types of human malignant melanoma cell lines revealed by expression patterns of mitochondrial and survival-apoptosis genes: implications for malignant melanoma therapy. Mol Cancer Ther. 2009; 8: 1292–1304.
- 51 Krejci P, Pejchalova K, Rosenbloom BE, et al. The antiapoptotic protein Api5 and its partner, high molecular weight FGF2, are up-regulated in B cell chronic lymphoid leukemia. J Leukoc Biol. 2007; 82: 1363–1364.
- 52 Issa JP, Ahuja N, Toyota M, et al. Accelerated age-related CpG island methylation in ulcerative colitis. Cancer Res. 2001; 61: 3573–3577.
- 53 Ozaki K, Nagasaka T, Notohara K, et al. Heterogeneous microsatellite instability observed within epithelium of ulcerative colitis. Int J Cancer. 2006; 119: 2513–2519.
- 54 Lashner BA, Shapiro BD, Husain A, et al. Evaluation of the usefulness of testing for p53 mutations in colorectal cancer surveillance for ulcerative colitis. Am J Gastroenterol. 1999; 94: 456–462.
- 55 Nathanson JW, Yadron NE, Farnan J, et al. p53 mutations are associated with dysplasia and progression of dysplasia in patients with Crohn's disease. Dig Dis Sci. 2008; 53: 474–480.
- 56 Sachdeva M, Zhu S, Wu F, et al. p53 represses c-Myc through induction of the tumor suppressor miR-145. Proc Natl Acad Sci U S A. 2009; 106: 3207–3212.
- 57 Olsen J, Gerds TA, Seidelin JB, et al. Diagnosis of ulcerative colitis before onset of inflammation by multivariate modeling of genome-wide gene expression data. Inflamm Bowel Dis. 2009; 15: 1032–1038.
- 58 Takagi T, Naito Y, Mizushima K, et al. Increased expression of microRNA in the inflamed colonic mucosa of patients with active ulcerative colitis. J Gastroenterol Hepatol. 2010; 25 Suppl 1: S129–133.
