title: Development of a high-resolution melting method for the screening of TNFAIP3 gene mutations.
abstract: Tumor necrosis factor, α-induced protein 3 (TNFAIP3) which encodes a ubiquitin-modifying enzyme (A20), acts as a negative regulator of the NF-κB pathway, and in lymphoma and autoimmune diseases it is frequently inactivated by mutations and/or deletions. We investigated the prevalence of the inactivation of TNFAIP3 in oral squamous cell carcinoma (OSCC). DNA was extracted from 81 cases of OSCC and 50 peripheral blood samples from normal controls. A high-resolution melting (HRM) analysis was used to characterize TNFAIP3 mutations, and the results were confirmed by direct DNA sequencing. Three mutations and three single-nucleotide polymorphisms (SNPs) were found to be associated with OSCC; the TNFAIP3 mutation occurred in 3.7% (3/81) of the OSCC cases examined. All mutations were in exon 7 [c.1081G>A (p.E361K), c.1398C>G (p.S466R) (rs200878487) and c.1760C>T (p.P587L) (rs150056192)], and p.E361K was identified as a novel mutation. We further used SIFT and PolyPhen-2 software to assess potentially functional mutations. Two SNPs, c.296‑20_296-18delCTC (rs71670547) and c.380T>G (p.F127C) (rs2230926), were located in exon 3, and c.2140C>T (p.P714S) was located in exon 9. A novel SNP, p.P714S differed from the one reported previously (p.P714A) (rs369155845) at that site. We also identified five SNPs in 50 normal Taiwanese individuals, and two of them [c.296‑15C>T (rs377482653) and c.305A>G (p.N102S) (rs146534657)] were not found in our OSCC tissue. HRM facilitated the screening of genetic changes. In addition, our results indicate that the prevalence of the TNFAIP3 mutation is low in OSCC.
authors.affiliation: Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, R.O.C.
authors.forename: Yuli Christine
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Epigenome Research Center, China Medical University Hospital, Taichung, Taiwan, R.O.C.
authors.forename: Ya-Sian
authors.initials: YS
authors.lastname: Chang
authors.affiliation: Division of Chest Medicine, Department of Internal Medicine, Changhua Christian Hospital, Changhua, Taiwan, R.O.C.
authors.forename: Chun-Chi
authors.initials: CC
authors.lastname: Chang
authors.affiliation: Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, R.O.C.
authors.forename: Ta-Chih
authors.initials: TC
authors.lastname: Liu
authors.affiliation: Environment-Omics‑Disease Research Center, China Medical University Hospital, Taichung, Taiwan, R.O.C.
authors.forename: Ying-Chin
authors.initials: YC
authors.lastname: Ko
authors.affiliation: Epigenome Research Center, China Medical University Hospital, Taichung, Taiwan, R.O.C.
authors.forename: Chien-Chin
authors.initials: CC
authors.lastname: Lee
authors.affiliation: Department of Kinesiology, Health and Leisure Studies, National University of Kaohsiung, Kaohsiung, Taiwan, R.O.C.
authors.forename: Shun-Jen
authors.initials: SJ
authors.lastname: Chang
authors.affiliation: Epigenome Research Center, China Medical University Hospital, Taichung, Taiwan, R.O.C.
authors.forename: Jan-Gowth
authors.initials: JG
authors.lastname: Chang
mesh_terms: D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D016022:Case-Control Studies; D004252:DNA Mutational Analysis / Q000379:methods; D005091:Exons; D005787:Gene Frequency; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D006239:Haplotypes; D006801:Humans; D009062:Mouth Neoplasms / Q000235:genetics*; D020125:Mutation, Missense; D011110:Polymorphism, Genetic; D060888:Real-Time Polymerase Chain Reaction; D017384:Sequence Deletion; D044366:Transition Temperature; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3 / Q000235:genetics*
chemical_list: C076529:TNFAIP3 protein, human; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3