Application Of High-Throughput Sequencing And Bioinformatics Analysis In Neurofibromatosis Type 1

Objection To explore the relationship between mutant genotypesphenotypes correlation in NF1 patients,and to find new pathogenic genes and their biological functions in diseases through bioinformatics analysis.Methods 1.A total of 122 patients with type 1 neurofibromatosis who were treated at the Ninth People’s Hospital of Shanghai Jiao Tong University School of Medicine from September 2016 to December 2018 were collected,and the mutation positions of NF1,NF2,and SPRED1 were detected by high-throughput sequencing.Points,and by combining its clinical manifestations and pedigrees,the gene-phenotype association analysis was performed.2.Screen differentially expressed genes through NF1-related gene expression data set(GSE14038)and gene methylation data set(E-GEOD21714),and combine GO and KEGG enrichment analysis,protein mole interaction analysis,and identify possible And the use of tissue samples for testing.3.For the validated pathogenic genes,use tumor primary cell culture technology and siRNA knockdown technology to silence the pathogenic genes,and combine CCK8 and transwell chambers to detect the proliferation,migration and Change in invasive ability.Results: 1.Through high-throughput analysis of peripheral blood in 122 patients with NF1,pathogenic mutations in 110 cases of NF1 were found in the study.Gene mutations in NF2 and SPRED1 were not found.No obvious hot spots were found in the distribution of mutation sites.There was no significant correlation between locus and phenotype.2.By analyzing the NF1-related data sets GSE14038 and E-GEOD21714,it was found that multiple genes including IGFBP1 were overexpressed in the NF1 patient group.By performing a subpopulation analysis on tissue samples,IGFBP1 was found in NF1-related benign tumor plexiform nerve Fibroids(PNF)and malignant tumor malignant peripheral neurofibrosphingoma(MPNST)are increasing.3.Using siRNA technology in PNF and MPNST primary Schwann cells to silence the expression of IGFBP1 gene,and found that it can inhibit the proliferation of PNF and MPNST cells,and inhibit the migration and invasion of MPNST cells.Conclusion: This study found that the NF1 gene mutation site in NF1 patients has no obvious mutation hot zone,and the genetic-phenotype association of the mutation site is not strong.It is necessary to find other related pathogenic genes to explain the complex clinical symptoms.Bioinformatics analysis found that IGFBP1 is a differentially expressed gene in NF1,and verified through histological and cytological levels that it may be a causative gene involved in the occurrence and malignancy of neurofibromas.