The Screening And Bioinformatics Analysing On Susceptibility Genes And Driving Genes Of A Rare Concurrent Cancer Family

Objective This research was intended to detect the susceptibility genes and driving genes in a pedigree diagnosed as concurrent cancer family and analyse the pathogenicity of the candidate genes.We sought to find somatic mutation sites and susceptibility genes mutation sites of this concurrent cancer.Methods(1)Clinical data and family pedigee were collected and the pedigree map was drawn.Tissue specimens and blood specimens were collected.(2)Genomic DNA was extracted with DNA Kit.Exome sequencing was completed with illumina hiseq 2500 plat.(3)Varscan2 was adopted for somatic mutation genes screening.The Picard and Genome Analysis Toolkit(GATK)methods were adopted for susceptibility genes screening.With data analysis and interpretation,we obtained the candidate genes.(4)The information of candidate genes whether reported or not were filtered in clinvar,HGMD,OMIM,Pubmed and NCBI.SIFT,Polyphen2,FATHMM and Mutationtaster were used to predict the risk of pathogenicity of the gene mutation sites.We used Oncomine database to analyze the expression of the mutant genes,c Bio Portal database to analyze mutation rate of candidate genes.The conservative analysis of somatic mutation sites were known by using the GERP++ and Phylop software.Results(1)The pedigree analysis showed an autosomal dominant inheritance.(2)We identified NDUFS7,the somatic mutation chr19:1393289(G>C)(p.R168S)that never reported in multiple gene database,as the only somatic mutations candidate gene.(3)We identified totally 17 non-synonymous germline mutations in 16 genes in peripheral blood samples of the concurrent cancer patients and one control person,including PGLYRP4,PEAR1,COL6A3,KIF1 A,ZNF717,ZNF141,SSPO,EPPK1,ZDHHC21,CNTRL,ASCC1,OR8U1,PABPC3,NPIPB6,RAB36 and CCDC117.(4)NDUFS7(p.R168S)mutation site was located in conserved sequences.SIFT,Polyphen2,FATHMM,Mutationtaster and other software predicted that site was probably pathogenic.(5)We screened two susceptible genes,ASCC1(c.G14A:p.R5H)and RAB36(c.G241A:p.D81N)which were predicted to be pathogenic by SIFT,Polyphen2,FATHMM and Mutationtaster software.Conclusion(1)This study collected a rare case of a three-generation concurrent cancer family.The pedigree analysis show an autosomal dominant inheritance.(2)Through whole exome sequencing,we selected a somatic mutation gene NDUFS7(p.R168S)and two susceptibility genes ASCC1(c.G14A:p.R5H)and RAB36(c.G241A:p.D81N)sites.(3)It is the first time to reveal that NDUFS7 may be the concurrent cancer pedigree driving gene,and ASCC1 and RAB36 may be susceptibility genes to concurrent cancer.