Characterization Of Cancer-related Genes Using Next Generation Sequencing And The Functional Study Of UNC5D Gene

Lung cancer is one of the most common cancers and the leading cause of cancer-related deaths worldwide.Non-small-cell lung cancers(NSCLCs),the most common lung cancers,are known to have complicated pathological features,which increase difficulties in NSCLC diagnosis and treatment.Thyroid carcinoma,the most frequent primary malignancy in the clinical endocrine organs,has a rapidly increasing incidence and prevalence in recent decades.Originating from thyroid follicular cells,papillary thyroid carcinoma(PTC)accounts for the vast majority of thyroid cancers(80%).Next generation sequencing technologies,featured with relatively low costs,high efficiency and high throughput,have been widely used in studies related to molecular diagnosis and target therapies in varies of cancers..Recently,the advent of next-generation sequencing(NGS)has dramatically revolutionized the molecular knowledge of NSCLC cancer by identifying lots of the pathogenic driver genes in large scale studies,including EGFR,KRAS,ALK,BRAF,PIK3 CA,MET,ERBB2,MAP2K1,NRAS,AKT1,etc..NSCLCs have already revealed genetically altered therapeutic targets such as EGFR inhibitors.The risk of cancer in thyroid nodules was estimated by the Bethesda system to be 5–15%.How to refine the assessment of cancer risk in thyroid nodules and to improve management of these patients remain to be a hotspot research program in thyroid fields.Recent studies using next-generation sequencing have indicated that thyroid carcinoma was predominantly driven by genetic and epigenetic alterations.Previous studies showed that a major class of driver genes of PTC were involved in the point mutations of BRAF,HRAS,KRAS,NRAS and fusion genes including RET/PTC etc..We performed next-generation sequencing combined with the function validation of mutant genes from three aspects: 1)We identified somatic gene mutations in 13 people with NSCLC by whole-exome/genome sequencing and confirmed them in an extended validation group of 88 people by targeted sequencing to clarify the driver genes and the mutation spectrum of Chinese NSCLC patients;2)We investigated the tumor-suppressive function of UNC5 D and examined whether identified UNC5 D point mutations in NSCLC cell lines disrupt the function of Unc5 D in tumor suppression;3)We tested whether the most complete next-generation sequencing(NGS)panel of genetic markers could significantly improve cancer diagnosis in thyroid nodules.The evaluation of 123 thyroid cancer samples using the targeted multi-PCR panel,which simultaneously tests for point mutations in 19 genes and for 8 types of gene fusions that occur in thyroid cancer.Taken together,our study has been investigating functional roles that UNC5 D plays in NSCLC and PTC etiology,expending the driver mutations spectrum of NSCLC and thyroid cancer,as well as providing insight into the potential candidates for diagnoses and targeted therapy in treating NSCLC and thyroid cancers in China.