Abstract
The retinoid acid receptor-β (RARβ) gene is one of the tumor suppressor genes (TSGs), which is frequently deleted or epigenetically silenced at an early stage of tumor progression. In this study we investigated the promoter methylation and expression status of the RARß gene in 60 surgically resected non-small cell lung cancer (NSCLC) tissue samples and 60 corresponding unchanged lung tissue samples, using methylation-specific PCR and real-time-polymerase chain reaction (qPCR) techniques. We correlated the results with the pathological features of tumors and clinical characteristics of patients. qPCR analysis detected a significantly lower RARß expression in the patients with adenocarcinoma (AC) and large cell carcinoma (LCC) than in those with squamous cell carcinoma (SCC) (AC vs. SCC, p = 0.032; AC and LCC vs. SCC, p = 0.013). Additionally, significantly lower expression of the RARß gene was revealed in the patients with non-squamous cell cancer with a history of smoking assessed as pack-years (PY <40 vs. PY ≥40, p = 0.045). Regarding RARß promoter methylation, we found significant differences in the methylation index in the SCC group when considering pTNM staging; with higher index values in T1a + T1b compared with T2a + T2b and T3 + T4 groups (p = 0.024). There was no correlation between the methylation status and expression level of the RARß gene, which suggests that other molecular mechanisms influence the RARß expression in NSCLC patients. In conclusion, different expression of the RARß gene in SCC and NSCC makes the RARß gene a valuable diagnostic marker for differentiating the NSCLC subtypes.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Agudo A, Bonet C, Travier N, González CA, Vineis P, Bueno-de-Mesquita HB, Trichopoulos D et al (2012) Impact of cigarette smoking on cancer risk in the European prospective investigation into cancer and nutrition study. J Clin Oncol 30:4550–4557
AJCC – American Joint Committee on Cancer Staging according to the IASCLC Staging Project (2010) 7th edn, Cancer
Bhagat R, Kumar SS, Vaderhobli S, Premalata CS, Pallavi VR, Ramesh G, Krishnamoorthy L (2014) Epigenetic alteration of p16 and retinoic acid receptor beta genes in the development of epithelial ovarian carcinoma. Tumour Biol 35(9):9069–9078
Brabender J, Metzger R, Salonga D, Danenberg KD, Danenberg PV, Hölscher AH, Schneider PM (2005) Comprehensive expression analysis of retinoic acid receptors and retinoid X receptors in non-small cell lung cancer implications for tumor development and prognosis. Carcinogenesis 26(3):525–530
Eissa S, Zohny SF, Shehata HH, Hegazy MG, Salem AM, Esmat M (2012) Urinary retinoic acid receptor-ß2 gene promoter methylation and hyaluronidase activity as noninvasive tests for diagnosis of bladder cancer. Clin Biochem 45(6):402–407
Ettinger DS, Akerley W, Bepler G, Blum MG, Chang A, Cheney RT, Chirieac LR et al (2010) Non-small cell lung cancer. J Natl Compr Cancer Netw 8(7):740–801
Feltus FA, Lee EK, Costello JF, Plass C, Vertino PM (2003) Predicting aberrant CpG island methylation. Proc Natl Acad Sci U S A 100:12253–12258
Gao T, He B, Pan Y, Li R, Xu Y, Chen L, Nie Z, Gu L, Wang S (2013) The association of retinoic acid receptor beta2 (RARß2) methylation status and prostate cancer risk: a systematic review and meta-analysis. PLoS One 8(5):e62950
Goldstraw P, Crowley J, Chansky K, Giroux DJ, Groome PA, Rami-Porta R, Postmus PE, Rusch V, Sobin L, International Association for the Study of Lung Cancer International Staging Committee, Participating Institutions (2007) The IASLC Lung Cancer Staging Project: proposals for the revision of the TNM stage groupings in the forthcoming (seventh) edition of the TNM Classification of malignant tumours. J Thorac Oncol 2:706–714
Greenlee RT, Murray T, Bolden S, Wingo PA (2000) Cancer statistics. CA Cancer J Clin 50:7–33
Khodyrev DS, Pronina IV, Rykov SV, Beresneva EV, Fridman MV, Kazubskaia TP, Loginov VI, Braga ÉA (2012) Methylation of some miRNA genes is involved in the regulation of their target genes RAR-beta2 and NKIRAS1 expression in lung cancer. Mol Biol (Mosk) 46(5):773–785
Li W, Deng J, Jiang P, Zeng X, Hu S, Tang J (2012) Methylation of the RASSF1A and RARß genes as a candidate biomarker for lung cancer. Exp Ther Med 3(6):1067–1071
Liu Z, Zhao J, Chen XF, Li W, Liu R, Lei Z, Liu X, Peng X, Xu K, Chen J, Liu H, Zhou QH, Zhang HT (2008) CpG island methylator phenotype involving tumor suppressor genes located on chromosome 3p in non-small cell lung cancer. Lung Cancer 62(1):15–22
Liu F, Killian JK, Yang M, Walker RL, Hong JA, Zhang M, Davis S, Zhang Y, Hussain M, Xi S, Rao M, Meltzer PA, Schrump DS (2010) Epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate. Oncogene 29(25):3650–3664
Nervi C, Grignani F (2014) RARs and microRNAs. Subcell Biochem 70:151–179
Pastuszak-Lewandoska D, Czarnecka KH, Migdalska-Sęk M, Nawrot E, Domańska D, Kiszałkiewicz J, Kordiak J, Antczak A, Górski P, Brzeziańska-Lasota E (2015) Decreased FAM107A expression in patients with non-small cell lung cancer. Adv Exp Med Biol 11:39–48
Picard E, Seguin C, Monhoven N, Rochette-Egly C, Siat J, Borrelly J, Martinet Y, Martinet N, Vignaud JM (1999) Expression of retinoid receptor genes and proteins in non-small-cell lung cancer. J Natl Cancer Inst 91(12):1059–1066
Sasco AJ, Secretan MB, Straif K (2004) Tobacco smoking and cancer: a brief review of recent epidemiological evidence. Lung Cancer 45(Suppl 2):3–9
Scesnaite A, Jarmalaite S, Mutanen P, Anttila S, Nyberg F, Benhamou S, Boffetta P, Husgafvel-Pursiainen K (2012) Similar DNA methylation pattern in lung tumours from smokers and never-smokers with second-hand tobacco smoke exposure. Mutagenesis 27(4):423–429
Sirchia SM, Ren M, Pili R (2002) Endogenous reactivation of the RARß2 tumor suppressor gene epigenetically silenced in breast cancer. Cancer Res 62:2455–2461
Souto-Garcia A, Fernández-Somoano A, Pascual T, Álvarez-Avellón SM, Tardón A (2012) Association of p21 Ser31Arg and p53Arg77Pro polymorphisms with lung cancer risk in CAPUA study. Lung Cancer Targets Ther 3:69–78
Staaf J, Isaksson S, Karlsson A, Jönsson M, Johansson L, Jönsson P, Botling J, Micke P, Baldetorp B, Planck M (2013) Landscape of somatic allelic imbalances and copy number alterations in human lung carcinoma. Int J Cancer 132(9):2020–2031
Tan C, Jin YT, Xu HY, Zhang CY, Zhang H, Zhang WM, Chen CM, Sun XY (2012) Correlation between RARbeta gene promoter methylation and P53 gene mutations in non-small cell lung cancer. Zhonghua Yi Xue Yi Chuan Xue Za Zhi 29(2):131–136
Virmani AK, Rathi A, Zochbauer-Muller S (2000) Promoter methylation and silencing of the retinoic acid receptor- beta gene in lung carcinomas. J Natl Cancer Inst (Bethesda) 92:1303–1307
Word B, Lyn-Cook LE, Mwamba B, Wang H, Lyn-Cook B, Hammons G (2013) Cigarette smoke condensate induces differential expression and promoter methylation profiles of critical genes involved in lung cancer in NL-20 lung cells in vitro: short-term and chronic exposure. Int J Toxicol 32(1):23–31
Zhang Y, Wang R, Song H, Huang G, Yi J, Zheng Y, Wang J, Chen L (2011) Methylation of multiple genes as a candidate biomarker in non-small cell lung cancer. Cancer Lett 303(1):21–28
Zhao Y, Zhou H, Ma K, Sun J, Feng X, Geng J, Gu J, Wang W, Zhang H, He Y, Guo S, Zhou X, Yu J, Lin Q (2013) Abnormal methylation of seven genes and their associations with clinical characteristics in early stage non-small cell lung cancer. Oncol Lett 5(4):1211–1218
Acknowledgements
This work was supported by a grant for young scientists of Medical University of Lodz no. 502-03/1-151-03/502-14-142.
Conflicts of Interest
The authors have no conflicts of interest concerning this study.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer International Publishing Switzerland
About this chapter
Cite this chapter
Dutkowska, A. et al. (2015). RARβ Promoter Methylation as an Epigenetic Mechanism of Gene Silencing in Non-small Cell Lung Cancer. In: Pokorski, M. (eds) Advances in Clinical Science. Advances in Experimental Medicine and Biology(), vol 878. Springer, Cham. https://doi.org/10.1007/5584_2015_159
Download citation
DOI: https://doi.org/10.1007/5584_2015_159
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-21496-2
Online ISBN: 978-3-319-21497-9
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)