Contents lists available at ScienceDirect Cytokine journal homepage: www.elsevier.com/locate/cytokine Genetic variations of IL-10: Identification of novel variations and evaluation of the impact of the SNPs/haplotype in the promoter region with the progression of Oral Squamous Cell Carcinoma in Indian population Upma Sharma a,j , Pallavi Singhal a , Kapil Bandil a , Anoop Kumar a,b , Surojit Bose c , Puneet Ahuja d , Molushree Kohli d , Ajay Kumar Dewan e , Juhi Tayal f , Basu Dev Banerjee g , Aditya Prasheri h , Ravi Mehrotra i , Veena Sharma j , Mausumi Bharadwaj a, ⁎ a Division of Molecular Genetics & Biochemistry, National Institute of Cancer Prevention & Research (NICPR-ICMR), I-7, Sector-39, Noida, India b National Institute of Biologicals, Noida, Uttar Pradesh, India c Barasat Cancer Hospital, Kolkata, India d Department of Oral Pathology, I.T.S. Dental College & Research, Greater Noida, India e Department of Surgical Oncology, Rajiv Gandhi Cancer Institute & Research Centre, New Delhi, India f Department of Research, Rajiv Gandhi Cancer Institute & Research Centre, New Delhi, India g Department of Biochemistry, University College of Medical Sciences, University of Delhi, Dilshad Garden, Delhi, India h Division of Molecular Immunology, National Institute of Cancer Prevention & Research (NICPR-ICMR), I-7, Sector-39, Noida, India i Division of Cytopathology, National Institute of Cancer Prevention & Research (NICPR-ICMR), I-7, Sector-39, Noida, India j Department of Bioscience and Biotechnology, Banasthali University, Rajasthan, India ARTICLE INFO Keywords: Interleukin 10 (IL-10) Oral Squamous Cell Carcinoma (OSCC) Denaturing High-Performance Liquid Chromatography (dHPLC) Amplification Refractory Mutation System- Polymerase Chain Reaction (ARMS-PCR) ABSTRACT The correlation of interleukin 10 (IL-10) with the outbreak and progression of cancer has been well established as it contributes to tumor immune evasion. Convincing number of evidences has been accumulated to reflect the critical correlation between IL-10 polymorphism and tumorogenesis. Several polymorphic sites at promoter regions have been reported to be associated with cancer susceptibility. The purpose of this study was to examine the effect of modulated genotypes in the promoter region of IL-10 gene with life-style habits in oral squamous cell carcinoma (OSCC) in the Indian population. A total of 300 subjects (100 OSCC, 50 precancer and 150 healthy controls) were recruited in this study. The IL-10 promoter region was amplified in 14 overlapping fragments by PCR and further screened through the high throughput technique of denaturing high-performance liquid chromatography (dHPLC) followed by sequencing. We identified three novel variations at positions (-924, -1045 & -1066); we also found some known SNPs (-592C/A, -657G/A, -851G/A, -819C/T, -1082A/G). The identified novel variations were submitted to the NCBI Gene Bank (accession numbers KT153594, KT291742 and KT291743). We also noticed a significant association of polymorphisms (-592C/A, -819C/T and -1082A/G) individually as well as in combination (haplotypes) along with lifestyle habits for the risk of oral carcinoma (p < 0.0001). We have reported three novel SNPs in the Indian population for the first time, and these SNPs may be associated with OSCC. Besides, we showed the first evidence of IL-10 haplotypes, i.e., CCG and CTG, may act as a biomarker for early detection of oral pre-cancerous/cancerous lesions or treatment management of oral carcinoma. 1. Introduction Cancer of oral cavity is a life-threatening disease with markedly increasing morbidity and mortality rates all over the world. It is ranked 11th globally, but it is the second-most disease causing mortality in developing nations like India [1]. In India, the majority cases of oral cancer consist of oral squamous cell carcinomas (OSCC), which arises from the epithelium of the oral mucosa [2,3]. The advent of Western lifestyle practices like cigarette smoking, alcohol consumption and chewing of smokeless tobacco products like khaini, gutka, supari, paan, etc. are possible causative factors of oral cancer in Indian population [4–6]. Any form of tobacco, either smoked or smokeless, can cause a wide spectrum of precancerous lesions or conditions like leukoplakia, erythroplakia, and oral submucous fibrosis (OSMF) that significantly http://dx.doi.org/10.1016/j.cyto.2017.09.016 Received 13 December 2016; Received in revised form 11 September 2017; Accepted 13 September 2017 ⁎ Corresponding author at: Division of Molecular Genetics & Biochemistry, National Institute of Cancer Prevention & Research (NICPR-ICMR), I-7, Sector 39, Noida 201301, India. E-mail addresses: bharadwajm@icmr.org.in, mausumi.bharadwaj@gmail.com (M. Bharadwaj). Cytokine xxx (xxxx) xxx–xxx 1043-4666/ © 2017 Published by Elsevier Ltd. Please cite this article as: Sharma, U., Cytokine (2017), http://dx.doi.org/10.1016/j.cyto.2017.09.016