Investigation of FOXP3 genetic variations at positions -2383 C/T and IVS9+459 T/C in southern Iranian patients with lung carcinoma

Document Type : Original Article


1 Cancer Immunology Group, Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

2 Department of Internal Medicine, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

3 Molecular Medicine Group, Graduate School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran


Objective(s): FOXP3 gene is an X-linked gene that encodes FOXP3 protein, an essential transcription factor in CD4+CD25+FOXP3+ regulatory T (Treg) cells.  We aimed, in the present study, to investigate the association of two FOXP3 polymorphisms, -2383 C/T (rs3761549) and IVS9+459 T/C (rs2280883), with lung cancer.
Materials and Methods:  In a case-control study we analyzed genotypes and alleles frequencies at -2383 C/T and IVS9+459 T/C positions in 156 patients with lung cancer and 156 age and sex matched healthy controls in Southern Iranian population, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. The data were verified by direct automated DNA sequencing.
Results: The frequency of -2383 T allele was significantly higher in the patients than in the control group (11.8% versus 5.9%, P-value=0.04, OR=2.13, 95%CI=1.04-4.54). T allele frequency at IVS9+459 T/C position was higher, compared to the controls, in the patients who presented the disease over 55 years old (69.9% versus 59.1%, P-value=0.04, OR=1.61, 95%CI=1.01-2.55) and also in SCLC patients (77.8% versus 59.1%, P-value=0.03, OR=2.42, 95%CI=1.05-5.59). No significant differences were found in the genotypes and haplotypes distributions between the cases and controls. A high degree of linkage disequilibrium was observed between two polymorphisms. 
Conclusion: As the first study dealing with -2383 C/T and IVS9+459 T/C in lung cancer, our data conclusively suggest the association of -2383 T allele with susceptibility to lung cancer in Iranian population. The association of IVS9+459 T allele with susceptibility to lung cancer in old patients suggests the age-dependent effects of FOXP3 gene on cancer occurrence.


1.  Fauci AS, kasper DL. Principles of Harrison's Internal Medicine. New York: McGraw-Hill; 2008.
2.  Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor Foxp3. Science 2003; 299:1057-1061.
3. Fontenot JD, Gavin MA, Rudensky AY. Foxp3 prog-rams the development and function of CD4+CD25+ regulatory T cells. Nat Immunol 2003; 4:330-336.
4.  Dejaco C, Duftner C, Grubeck-Loebenstein B, Schirmer M. Imbalance of regulatory T cells in human autoimmune diseases. Immunology 2006; 117:289-300.
5. Von Boehmer H. Mechanisms of suppression by suppressor T cells. Nat Immunol 2005; 6:338-344.
6.  Ishibashi Y, Tanaka S, Tajima K, Yoshida T, Kuwano H. Expression of Foxp3 in non-small cell lung cancer patients is significantly higher in tumor tissues than in normal tissues, especially in tumors smaller than 30 mm. Oncol Rep 2006; 15:1315-1319.
7.  Zhang HY, Sun H. Up-regulation of Foxp3 inhibits cell proliferation, migration and invasion in epithelial ovarian cancer. Cancer Lett. 2010; 287:91-97.
8.  Zeng C, Yao Y, Jie W, Zhang M, Hu X, Zhao Y, et al. Up-regulation of Foxp3 participates in progression of cervical cancer. Cancer Immunol Immunother 2013; 62:481-487.
9.  Dimitrakopoulos FI, Papadaki H, Antonacopoulou AG, Kottorou A, Gotsis AD, Scopa C, et al. Association of FOXP3 expression with non-small cell lung cancer. Anticancer Res 2011; 31:1677-1683.
10. Zuo T, Wang L, Morrison C, Chang X, Zhang H, Li W, et al. FOXP3 is an X-linked breast cancer suppressor gene and an important repressor of the HER-2/ErbB2 oncogene. Cell 2007; 129:1275-1286.
11. Wang L, Liu R, Ribick M, Zheng P, Liu Y. FOXP3 as an X-linked tumor suppressor. Discov Med 2010; 10:322-328.
12. Li W, Wang L, Katoh H, Liu R, Zheng P, Liu Y. Identification of a tumor suppressor relay between the FOXP3 and the Hippo pathways in breast and prostate cancers. Cancer Res 2011; 71:2162-2171.
13. Wang L, Liu R, Li W, Chen C, Katoh H, Chen GY, et al. Somatic single hits inactivate the X-linked tumor suppressor FOXP3 in the prostate. Cancer Cell 2009; 16:336-346.
14. Zuo T, Liu R, Zhang H, Chang X, Liu Y, Wang L, et al. FOXP3 is a novel transcriptional repressor for the breast cancer oncogene SKP2. J Clin Invest 2007; 117:3765-3773.
15. Raskin L, Rennert G, Gruber SB. FOXP3 germline polymorphisms are not associated with risk of breast cancer. Cancer Genet Cytogenet 2009; 190:40-42.
16. Mojtahedi Z, Erfani N, haghshenas MR, hoseini SV, Ghaderi A. Association of  Foxp3/Scurfin germline polymorphisms(C-2383T/rs3761549) with colorectal cancer. Ann Colorectal Res 2013; 1:12-16.
17. Zheng J, Deng J, Jiang L, Yang L, You Y, Hu M, et al. Heterozygous genetic variations of FOXP3 in Xp11.23 elevate breast cancer risk in Chinese population via skewed X-chromosome inactivation. Hum Mutat 2013; 34:619-628.
18. Chen Y, Zhang H, Liao W, Zhou J, He G, Xie X, et al. FOXP3 gene polymorphism is associated with hepatitis B-related hepatocellular carcinoma in China. J Exp Clin Cancer Res 2013; 32:39.
19. He YQ, Bo Q, Yong W, Qiu ZX, Li YL, Li WM. FoxP3 genetic variants and risk of non-small cell lung cancer in the Chinese Han population. Gene 2013; 531:422-425.
20. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988; 16:1215.
21. Owen CJ, Eden JA, Jennings CE, Wilson V, Cheetham TD, Pearce SH. Genetic association studies of the FOXP3 gene in Graves' disease and autoimmune Addison's disease in the United Kingdom population. J Mol Endocrinol 2006; 37:97-104.
22. Ban Y, Tozaki T, Tobe T, Jacobson EM, Concepcion ES, Tomer Y. The regulatory T cell gene FOXP3 and genetic susceptibility to thyroid autoimmunity: an association analysis in Caucasian and Japanese cohorts. J Autoimmun 2007; 28:201-207.
23. Gao L, Li K, Li F, Li H, Liu L, Wang L, et al. Polymorphisms in the FOXP3 gene in Han Chinese psoriasis patients. J Dermatol Sci 2010; 57:51-56.
24. Lan Y, Tang XS, Qin J, Wu J, Qin JM. [Association of transcription factor FOXP3 gene polymorphism with genetic susceptibility to systematic lupus erythematosus in Guangxi Zhuang population]. Zhonghua Yi Xue Yi Chuan Xue Za Zhi 2010; 27:433-436.
25. Lin YC, Lee JH, Wu AS, Tsai CY, Yu HH, Wang LC, et al. Association of single-nucleotide polymorphisms in FOXP3 gene with systemic lupus erythematosus susceptibility: a case-control study. Lupus 2011; 20:137-143.
26. Song QH, Shen Z, Xing XJ, Yin R, Wu YZ, You Y, et al. An association study of single nucleotide polymorphisms of the FOXP3 intron-1 and the risk of Psoriasis vulgaris. Indian J Biochem Biophys 2012; 49:25-35.
27. Zhang J, Chen Y, Jia G, Chen X, Lu J, Yang H, et al. FOXP3 -3279 and IVS9+459 polymorphisms are associated with genetic susceptibility to myasthenia gravis. Neurosci Lett 2013; 534:274-278.
28. Conteduca G, Rossi A, Megiorni F, Parodi A, Ferrera F, Tardito S, et al. Single nucleotide polymorphisms in the promoter regions of Foxp3 and ICOSLG genes are associated with Alopecia areata. Clin Exp Med 2014; 14:91-97.
29. Bryant AS, Cerfolio RJ. Differences in outcomes between younger and older patients with non-small cell lung cancer. Ann Thorac Surg 2008; 85:1735-1739.
30. Skarin AT, Herbst RS, Leong TL, Bailey A, Sugarbaker D. Lung cancer in patients under age 40. Lung Cancer 2001; 32:255-264.
31. Weiskopf D, Weinberger B, Grubeck-Loebenstein B. The aging of the immune system. Transpl Int 2009; 22:1041-1050.
32. Zanussi S, Serraino D, Dolcetti R, Berretta M, De Paoli P. Cancer, aging and immune reconstitution. Anticancer Agents Med Chem 2013; 13:1310-1324.
33. Shaw AC, Goldstein DR, Montgomery RR. Age-dependent dysregulation of innate immunity. Nat Rev Immunol 2013; 13:875-887.
34. Pan XD, Mao YQ, Zhu LJ, Li J, Xie Y, Wang L, et al. Changes of regulatory T cells and FoxP3 gene expression in the aging process and its relationship with lung tumors in humans and mice. Chin Med J (Engl) 2012; 125:2004-2011.
35. Nishioka T, Shimizu J, Iida R, Yamazaki S, Sakaguchi S. CD4+CD25+Foxp3+ T cells and CD4+CD25-Foxp3+ T cells in aged mice. J Immunol 2006; 176:6586-6593.
36. Gregg R, Smith CM, Clark FJ, Dunnion D,  Khan N, Chakraverty R, et al. The number of human peripheral blood CD4+ CD25high regulatory T cells increases with age. Clin Exp Immunol 2005; 140:540-546.
37. Kumar V, Abbas AK, J.C. a. Robbins and Cotran Pathologic Basis of Disease. 8th ed. Philadelphia: Saunders; 2012.
38. Inoue N, Watanabe M, Morita M, Tomizawa R, Akamizu T, Tatsumi K, et al. Association of functional polymorphisms related to the transcriptional level of FOXP3 with prognosis of autoimmune thyroid diseases. Clin Exp Immunol 2010; 162:402-406.
39. Andre GM, Barbosa CP, Teles JS, Vilarino FL, Christofolini DM, Bianco B. Analysis of FOXP3 polymorphisms in infertile women with and without endometriosis. Fertil Steril 2011; 95:2223-2227.
40. Park O, Grishina I, Leung PS, Gershwin ME, Prindiville T. Analysis of the Foxp3/scurfin gene in Crohn's disease. Ann N Y Acad Sci 2005; 1051:218-228.