Identification and association of the single nucleotide polymorphisms, C-509T, C+466T and T+869C, of the TGF-β1 gene in patients with asthma and their influence on the mRNA expression level of TGF-β1

Panek,Michał; Pietras,Tadeusz; Fabijan,Artur; Zioło,Jan; Wieteska,Łukasz; Małachowska,Beata; Fendler,Wojciech; Szemraj,Janusz; Kuna,Piotr

doi:10.3892/ijmm.2014.1894

Identification and association of the single nucleotide polymorphisms, C-509T, C+466T and T+869C, of the TGF-β1 gene in patients with asthma and their influence on the mRNA expression level of TGF-β1

Authors:
- Michał Panek
- Tadeusz Pietras
- Artur Fabijan
- Jan Zioło
- Łukasz Wieteska
- Beata Małachowska
- Wojciech Fendler
- Janusz Szemraj
- Piotr Kuna
View Affiliations

Affiliations: Department of Internal Medicine, Asthma and Allergy, Medical University of Lodz, 90-153 Lodz, Poland, Department of Pneumology and Allergology, Medical University of Lodz, 90-153 Lodz, Poland, Students Research Group, Department of Internal Medicine, Asthma and Allergy, Medical University of Lodz, 90-153 Lodz, Poland , Department of Medical Biochemistry, Medical University of Lodz, 92-215 Lodz, Poland, Department of Pediatrics, Oncology, Hematology and Diabetology, Medical University of Lodz, 91-738 Lodz, Poland
Published online on: August 11, 2014 https://doi.org/10.3892/ijmm.2014.1894
Pages: 975-986
Copyright: © Panek et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Transforming growth factor-β1 (TGF-β1) is an important fibrogenic and immunomodulatory cytokine participating in the pathogenesis of a number of illnesses related to the growth, differentiation and migration of cells. It also plays a key role in inflammation, atherosclerosis, vascular inflammation and asthma. The aim of the present study was to evaluate the association between the expression of the TGF-β1 gene and its genetic polymorphisms, and the disease phenotype. The study comprised 173 patients with asthma, as well as 163 healthy volunteers as a control group. The gender profiles of the groups were similar (p=0.8415). Genotyping was performed by polymerase chain reaction (PCR)-high resolution melting (HRM). The results were verified by sequencing. Gene expression was evaluated by RT-PCR. This study evaluated the role and frequency of genetic polymorphisms (C-509T, C+466T and T+869C) of the TGF-β1 gene in the study group (patients with asthma) and the control group (healthy volunteers). The results obtained for the patients and healthy controls were as follows: C-509T single nucleotide polymorphism (SNP) (controls, TT/CT/CC-0.4444/0.5309/0.0247; patients, TT/CT/CC-0.3699/0.6012/0.0289), C+466T SNP (controls, TT/CT/CC-1.000/0.000/0.000; patients, TT/CT/CC-1.000/0.000/0.000) and T+869C SNP (controls, TT/CT/CC-1.000/0.000/0.000; patients, TT/CT/CC-1.000/0.000/0.000). Only the C-509T polymorphism was found to play a significant role in the pathogenesis of asthma, as well as a risk factor in the loss of the clinical control of the disease [TT vs. CC/CT, odds ratio (OR) 2.38; confidence interval (CI) 1.22-4.66; p=0.0103]. A significant difference was noted between the study and control groups with regard to the mRNA expression of TGF-β1 (p=0.0133). A higher level of expression of the TGF-β1 gene correlated with the time of diagnosis of patients over 16 years of age (p=0.0255). This study demonstrates that the C-509T SNP is a significant clinical risk factor for asthma and that the TGF-β1 cytokine contributes to the progression of the illness.

View Figures

View References

1	Droszcz W and Grzanka A: Asthma in adolescents and adults. Wydawnictwo Lekarskie PZWL; Warszawa: 2010
2	Genuneit J, Cantelmo JL, Weinmayr G, Wong GW, Cooper PJ, Riikjärv MA, Gotua M, Kabesch M, von Mutius E, Forastiere F, Crane J, Nystad W, El-Sharif N, Batlles-Garrido J, García-Marcos L, García-Hernández G, Morales-Suarez-Varela M, Nilsson L, Bråbäck L, Saraçlar Y, Weiland SK, Cookson WO, Strachan D and Moffatt MF; ISAAC Phase 2 Study Group. A multi-centre study of candidate genes for wheeze and allergy: the International Study of Asthma and Allergies in Childhood Phase 2. Clin Exp Allergy. 39:1875–1888. 2009. View Article : Google Scholar : PubMed/NCBI
3	Anderson GP: Endotyping asthma: new insights into key pathogenic mechanisms in a complex, heterogeneous disease. Lancet. 372:1107–1119. 2008. View Article : Google Scholar : PubMed/NCBI
4	Djukanović R, Roche WR, Wilson JW, Beasley CR, Twentyman OP, Howarth RH and Holgate ST: Mucosal inflammation in asthma. Am Rev Respir Dis. 142:434–457. 1990.
5	Sumi Y and Hamid Q: Airway remodeling in asthma. Allergol Int. 56:341–348. 2007. View Article : Google Scholar
6	Romagnani S: The increased prevalence of allergy and the hygiene hypothesis: missing immune deviation, reduced immune suppression, or both? Immunology. 112:352–363. 2004. View Article : Google Scholar : PubMed/NCBI
7	Wang J and Young IG: Eosinophilic inflammation: mechanisms regulating IL-5 transcription in human T lymphocytes. Allergy. 62:1131–1138. 2007. View Article : Google Scholar : PubMed/NCBI
8	Yang YC, Zhang N, Van Crombruggen K, Hu GH, Hong SL and Bachert C: Transforming growth factor-beta1 in inflammatory airway disease: a key for understanding inflammation and remodeling. Allergy. 67:1193–1202. 2012. View Article : Google Scholar : PubMed/NCBI
9	Duvernelle C, Freund V and Frossard N: Transforming growth factor-beta and its role in asthma. Pulm Pharmacol Ther. 16:181–196. 2003. View Article : Google Scholar : PubMed/NCBI
10	Park HK, Park HW, Jeon SG, Shin ES, Gho YS, Cho SH, Kim YY and Kim YK: Distinct association of genetic variations of vascular endothelial growth factor, transforming growth factor-β, and fibroblast growth factor receptors with atopy and airway hyperresponsiveness. Allergy. 63:447–453. 2008.PubMed/NCBI
11	Attisano L and Wrana JL: Signal transduction by the TGF-beta superfamily. Science. 296:1646–1647. 2002. View Article : Google Scholar : PubMed/NCBI
12	Letterio JJ and Roberts AB: TGF-beta: a critical modulator of immune cell function. Clin Immunol Immunopathol. 84:244–250. 1997. View Article : Google Scholar : PubMed/NCBI
13	Ashcroft GS: Bidirectional regulation of macrophage function by TGF-beta. Microbes Infect. 1:1275–1282. 1999. View Article : Google Scholar : PubMed/NCBI
14	Pancewicz SA, Izycka A, Klibingat M, Zajkowska JM, Swierzbińska-Pijanowska R, Kondrusik M, Grygorczuk SS, Izycki T and Hermanowska-Szpakowicz T: Transforming growth factor beta, metalloproteinase 2 and its tissue inhibitor 2 in the serum from patients with early and late boreliosis. Pol Merkur Lekarski. 25:495–499. 2008.(In Polish).
15	Kim DH and Kim SJ: Transforming growth factor-beta receptors: Role in physiology and disease. J Biomed Sci. 3:143–158. 1996. View Article : Google Scholar : PubMed/NCBI
16	Attisano L and Wrana JL: Signal transduction by members of the transforming growth factor-beta superfamily. Cytokine Growth Factor Rev. 7:327–339. 1996. View Article : Google Scholar : PubMed/NCBI
17	Lagna G, Hata A, Hemmati-Brivanlou A and Massagué J: Partnership between DPC4 and SMAD proteins in TGF-beta signalling pathways. Nature. 383:832–836. 1996. View Article : Google Scholar : PubMed/NCBI
18	Wrana JL: Crossing Smads. Sci STKE. 14:2000. re12000.
19	Whitman M: Smads and early developmental signaling by the TGFbeta superfamily. Genes Dev. 12:2445–2462. 1998. View Article : Google Scholar : PubMed/NCBI
20	Heldin CH, Miyazono K and ten Dijke P: TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature. 390:465–471. 1997. View Article : Google Scholar : PubMed/NCBI
21	Kim SJ, Glick A, Sporn MB and Roberts AB: Characterization of the promoter region of the human transforming growth factor-beta 1 gene. J Biol Chem. 264:402–408. 1989.PubMed/NCBI
22	Kim SJ, Park K, Rudkin BB, Dey BR, Sporn MB and Roberts AB: Nerve growth factor induces transcription of transforming growth factor-beta 1 through a specific promoter element in PC12 cells. J Biol Chem. 269:3739–3744. 1994.PubMed/NCBI
23	Kim SJ, Angel P, Lafyatis R, Hattori K, Kim KY, Sporn MB, Karin M and Roberts AB: Autoinduction of transforming growth factor beta 1 is mediated by the AP-1 complex. Mol Cell Biol. 10:1492–1497. 1990.PubMed/NCBI
24	Kim SJ, Romeo D, Yoo YD and Park K: Transforming growth factor-beta: expression in normal and pathological conditions. Horm Res. 42:5–8. 1994. View Article : Google Scholar : PubMed/NCBI
25	Watanabe Y, Kinoshita A, Yamada T, Ohta T, Kishino T, Matsumoto N, Ishikawa M, Niikawa N and Yoshiura K: A catalog of 106 single-nucleotide polymorphisms (SNPs) and 11 other types of variations in genes for transforming growth factor-beta1 (TGF-beta1) and its signaling pathway. J Hum Genet. 47:478–483. 2002. View Article : Google Scholar : PubMed/NCBI
26	Mak JC, Leung HC, Ho SP, Law BK, Ho AS, Lam WK, Ip MS and Chan-Yeung MM: Analysis of TGF-beta(1) gene polymorphisms in Hong Kong Chinese patients with asthma. J Allergy Clin Immunol. 117:92–96. 2006. View Article : Google Scholar : PubMed/NCBI
27	Grainger DJ, Heathcote K, Chiano M, Snieder H, Kemp PR, Metcalfe JC, et al: Genetic control of the circulating concentration of transforming growth factor type beta1. Hum Mol Genet. 8:93–97. 1999. View Article : Google Scholar : PubMed/NCBI
28	Hobbs K, Negri J, Klinnert M, Rosenwasser LJ and Borish L: Interleukin-10 and transforming growth factor-beta promoter polymorphisms in allergies and asthma. Am J Respir Crit Care Med. 158:1958–1962. 1998. View Article : Google Scholar : PubMed/NCBI
29	Pulleyn LJ, Newton R, Adcock IM and Barnes PJ: TGFbeta1 allele association with asthma severity. Hum Genet. 109:623–627. 2001. View Article : Google Scholar : PubMed/NCBI
30	Silverman ES, Palmer LJ, Subramaniam V, Hallock A, Mathew S, Vallone J, et al: Transforming growth factor-beta1 promoter polymorphism C−509T is associated with asthma. Am J Respir Crit Care Med. 169:214–219. 2004.
31	Nagpal K, Sharma S, B-Rao C, Nahid S, Niphadkar PV, Sharma SK and Ghosh B: TGFbeta1 haplotypes and asthma in Indian populations. J Allergy Clin Immunol. 115:527–533. 2005. View Article : Google Scholar : PubMed/NCBI
32	Buckova D, Izakovicová Hollá L, Benes P, Znojil V and Vácha J: TGF-beta1 gene polymorphisms. Allergy. 56:1236–1237. 2001. View Article : Google Scholar : PubMed/NCBI
33	Heinzmann A, Bauer E, Ganter K, Kurz T and Deichmann KA: Polymorphisms of the TGF-beta1 gene are not associated with bronchial asthma in Caucasian children. Pediatr Allergy Immunol. 16:310–314. 2005. View Article : Google Scholar : PubMed/NCBI
34	National Center for Biotechnology Information. The U.S. National Institutes of Health. http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=200482214.
35	Panek M, Pietras T, Szemraj J, Fabijan A and Kuna P: Identification and association of TGFβ-1 expression in patients with asthma in a Polish population - Lodz metropolitan area study. Int J Biochem Mol Biol. 4:67–74. 2013.
36	Panek M, Pietras T, Fabijan A, Miłanowski M, Wieteska L, Górski P, Kuna P and Szemraj J: Effect of glucocorticoid receptor gene polymorphisms on asthma phenotypes. Exp Ther Med. 5:572–580. 2013.PubMed/NCBI
37	Panek M, Pietras T, Antczak A, Fabijan A, Przemęcka M, Górski P, Kuna P and Szemraj J: The N363S and I559N single nucleotide polymorphisms of the h-GR/NR3C1 gene in patients with bronchial asthma. Int J Mol Med. 30:142–150. 2012.PubMed/NCBI
38	Panek M, Pietras T, Antczak A, Górski P, Kuna P and Szemraj J: The role of functional single nucleotide polymorphisms of the human glucocorticoid receptor gene NR3C1 in Polish patients with bronchial asthma. Mol Biol Rep. 39:4749–4757. 2012. View Article : Google Scholar
39	Nathan RA, Sorkness CA, Kosinski M, Schatz M, Li JT, Marcus P, Murray JJ and Pendergraft TB: Development of the asthma control test: a survey for assessing asthma control. J Allergy Clin Immunol. 113:59–65. 2004. View Article : Google Scholar : PubMed/NCBI
40	Chomczynski P and Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 162:156–159. 1987. View Article : Google Scholar : PubMed/NCBI
41	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001.
42	Winer J, Jung CK, Shackel I and Williams PM: Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem. 270:41–49. 1999. View Article : Google Scholar
43	Li H, Romieu I, Wu H, Sienra-Monge JJ, Ramírez-Aguilar M, del Río-Navarro BE, del Lara-Sánchez IC, Kistner EO, Gjessing HK and London SJ: Genetic polymorphisms in transforming growth factor beta-1 (TGFB1) and childhood asthma and atopy. Hum Genet. 121:529–538. 2007. View Article : Google Scholar : PubMed/NCBI
44	Flanders KC and Burmester JK: Medical applications of transforming growth factor-beta. Clin Med Res. 1:13–20. 2003. View Article : Google Scholar : PubMed/NCBI
45	Horowitz JC, Lee DY, Waghray M, Keshamouni VG, Thomas PE, Zhang H, Cui Z and Thannickal VJ: Activation of the pro-survival phosphatidylinositol 3-kinase/AKT pathway by transforming growth factor-beta1 in mesenchymal cells is mediated by p38 MAPK-dependent induction of an autocrine growth factor. J Biol Chem. 279:1359–1367. 2004. View Article : Google Scholar : PubMed/NCBI
46	Lee KY and Bae SC: TGF-beta-dependent cell growth arrest and apoptosis. J Biochem Mol Biol. 35:47–53. 2002. View Article : Google Scholar : PubMed/NCBI
47	Li MO, Wan YY, Sanjabi S, Robertson AK and Flavell RA: Transforming growth factor-beta regulation of immune responses. Annu Rev Immunol. 24:99–146. 2006. View Article : Google Scholar : PubMed/NCBI
48	Stępień-Wyrobiec O, Hrycek A and Wyrobiec G: Transforming growth factor beta (TGF-beta): its structure, function, and role in the pathogenesis of systemic lupus erythematosus. Postepy Hig Med Dosw (Online). 62:688–693. 2008.(In Polish).
49	Bierie B and Moses HL: Tumour microenvironment: TGFbeta: the molecular Jekyll and Hyde of cancer. Nat Rev Cancer. 6:506–520. 2006. View Article : Google Scholar : PubMed/NCBI
50	Fleisch MC, Maxwell CA and Barcellos-Hoff MH: The pleiotropic roles of transforming growth factor beta in homeostasis and carciongenesis of endocrine organs. Endocr Relat Cancer. 13:379–400. 2006. View Article : Google Scholar : PubMed/NCBI
51	Niemczyk M, Foroncewicz B and Mucha K: The role of TGF beta. Pol Arch Med Wewn. 113:401–408. 2005.(In Polish).
52	Kim SG, Jong HS, Kim TY, Lee JW, Kim NK, Hong SH and Bang YJ: Transforming growth factor-beta 1 induces apoptosis through Fas ligand-independent activation of the Fas death pathway in human gastric SNU-620 carcinoma cells. Mol Biol Cell. 15:420–434. 2004. View Article : Google Scholar
53	Moustakas A, Souchelnytskyi S and Heldin CH: Smad regulation in TGF-beta signal transduction. J Cell Sci. 114:4359–4369. 2001.PubMed/NCBI