Study On Association Between Polymorphism Of TRAF2,TRAF3,NFKBIA,CHUK And MAP2K4 Genes And Nasopharyngeal Carcinoma Susceptibility In Chinese Han Population

Objective:Nasopharyngeal carcinoma(NPC)is an Epstein-Barr virus(EBV)associated malignancy.The EBV oncoprotein latent membrane protein 1(LMP1)is the primary oncogene of EBV infection through its signaling cascade and its connections to other pathways including NF-?B,and JNK signaling,which plays an important role in the pathogenesis of NPC.Our study is aim to investigate the association between single nucleotide polymorphisms(SNPs)within genes involved in NF-?B and JNK signaling pathways and risk of NPC in Chinese Han population.It may provide novel markers for the NPC early screening,and set up of prevention and intervention systems for high-risk population of NPC.Methods:350 NPC patients pathological diagnosed for the first time at Sichuan Cancer Hospital from January 2017 to September 2018 were recruited.580controls were recruited from physical examination center of Sichuan Provincial People's Hospital from July to September 2018.Candicated genes that related to the causation of NPC through the database KEGG,NCBI involved in the NF-?B and JNK signaling pathways were selected,literature survey concern to as sociation between candicate genes and causation of NPC were checked on OMIM.Altogether149 SNPs were covered by the 15 SNPs in TRAF2(rs7852970?rs3739942?rs5901110)?TRAF3(rs1131877?rs1051750?rs10133111?rs11160707)?NFKBIA(rs8904?rs2233406?rs2233409?rs145753299?rs2233407?rs17103265)?CHUK(rs2230804)andMAK2P4(rs4792219)wereseletedfromthe Ensemble,dbSNP,1000 Genomes Project and ExAC meet the following criteria:minor allele frequency(MAF)?10%in HCB(Chinese Han in Beijing)or East Asian(EAS);location within the coding region(non-synonymous SNPs),the 3'and5'untranslated regions(UTRs)and the promoter;linkage disequilibrium(LD,r~2?0.80)between the SNPs.DNA was extracted from peripheral blood leukocytes by using kit.Concentrations of all samples were higher than 20 ng/?L and 260/280value was between 1.7 and 2.0.Then,the qualified DNA samples were subjected to agarose gel electrophoresis,to confirm the DNA samples were qualified.15 selected SNPs were genotyped by using qualified samples(MassARRAY),and 5%samples were randomly selected to verify the genotyping results by Sanger sequencing.We used Haploview to calculate Hardy-Weinberg equilibrium(HWE),MAF of HCB and LD value between selected SNPs in controls.Odds ratios(ORs),95%confidence intervals(95%CIs)and p value for associations between genotypes and NPC risk were calculated by Plink1.03.Results:Genetic typing on TRAF2(rs7852970),TRAF3(rs1051750,rs11160707)and NFKBIA(rs145753299)were failed.We used rs17250694instead of TRAF2(rs7852970)(LD r~2>0.8),rs2075771 instead of TRAF3(rs1051750)and rs2273650 instead of NFKBIA(rs145753299).We did not find alternative sites for TRAF3(rs11160707).Except for NFKBIA(rs2233409),all MAF were?10%and the genotype distribution of all other genotyped polymorphisms was consistent with HWE in the control group(P>0.05).MAP2K4 allele rs4792219,after removing samples that failed for genotyping,there were 345 patients and 570 normal controls.The result showed HCB MAF=17.6%,there was a statistical difference between G/G-G/A and A/A group in recessive mode,OR=0.36,95%CI=0.12-1.07,P=0.044;the genotype frequencies of the remaining sites were no significant difference between the case group and the normal control group(P>0.05).Conclusion:Our data showed SNP in MAP2K4 gene might affect NPC risk,rs4792219 may be a useful biomarker for NPC early screening.