Study On The Molecular Markers Of Lymph Node Metastasis In Esophageal Squamous Cell Carcinoma

Esophageal squamous-cell carcinoma is the main pathological form of esophageal cancer patients in China,accounting for 95%of cases of esophageal cancer.Its morbidity and mortality rate account for the 9^th and 6^th place of malignant tumors.Radical surgical resection is the first choice for the treatment of esophageal cancer,but still facing high local recurrence rate and poor prognosis.Lymph node metastasis is one of the main factors for local recurrence of esophageal cancer.In recent years,molecular markers study related to lymph node metastasis using sequencing technology has become a hotspot in this research field.The molecular markers related to lymph node metastasis in esophageal cancer which discovered so far are mainly abnormality in gene expression level.Molecular markers related to lymph node metastasis in esophageal cancer at the gene mutation level are rarely reported.ObjectiveRegional lymph node metastasis is one of the main factors for recurrence of postoperative esophageal patient.The determination of the extent of lymph node dissection in the surgical plan requires a comprehensive consideration of the patient's surgical injury and the probability of cancer recurrence.Several molecular markers associated with esophageal cancer lymph node metastasis have been discovered at gene expression level so far.However,these markers are not sufficient to measure the risk level of lymph node metastasis of esophageal cancer.In order to explore new or potential molecular markers of lymph node metastasis of esophageal cancer and evaluate the risk of lymph node metastasis of esophageal cancer more accurately,We studied the whole exome somatic mutation data and the clinical data of patients with esophageal squamous-cell carcinoma.Statistical and bioinformatics methods were used to study the molecular markers of lymph node metastasis gene expression levels and their sensitivity and specificity.Exploring the functional distribution and molecular mechanism of molecular markers at gene mutation level provided a technical and theoretical basis for predicting the risk of lymph node metastasis of esophageal squamous-cell carcinoma in this study.MethodsTotally 91 patients with esophageal squamous-cell carcinoma were divided into two groups according to whether lymph node metastasis occurred.Inter-group difference analysis was performed on 17 indicators using single factor analysis.The correlation between the probability of lymph node metastasis and the difference between groups were analyzed by binary logistic regression.ROC curve analysis was used to evaluate the application value of significant related indicators.The Yoden's index method was used to judge the optimal cutoff value,sensitivity and specificity of potential molecular markers.The whole exome genes were divided into 7 different ranges according to the gene function category and gene mutation frequency.The gene set associated with the potential molecular markers was judged by the consistency of the prediction results in different ranges and result of the total exome.KEGG signaling pathway analysis and KOG orthologous sequence enrichment were used to study the distribution of potential molecular marker-related gene sets in different functional pathways.Results?1?Univariate analysis of differences between groups showed that the age of onset?t=2.087,P<0.05?and the GC%of the wild-type base in somatic mutation at the first codon position?GWSMFCP??t=2.542,P<0.05?were statistically significant difference between groups.?2?Binary logistic regression analysis showed that GWSMFCP was significantly negatively correlated with the probability of lymph node metastasis of esophageal squamous-cell carcinoma?P<0.05?,which was an independent risk factor for lymph node metastasis.?3?ROC curve analysis showed that GWSMFCP was the protective factor for lymph node metastasis of esophageal squamous-cell carcinoma.The max Youden's index was 0.277.The sensitivity and specificity of predicting lymph node metastasis risk were 56.6%and 71.1%,respectively.?4?Difference,correlation and predictive performance between GWSMFCP markers and lymph node metastasis in patients with esophageal squamous-cell carcinoma in the range of three exogenous genes,including all exomes,non-driven genes,and low-frequency mutant non-driven genes.The evaluation indicators were consistent,indicating that the mechanism of action of this indicator was in the widely distributed low-frequency mutant non-driven genes.?5?There was no difference,correlation and application value between GWSMFCP index and lymph node metastasis in patients with esophageal squamous-cell carcinoma or not in the range of high frequency mutant non-driven gene,driver gene,functional inactivation-driven gene,dysfunctional activation-driven gene and unclassified driver gene.The results indicate that the mechanism of this indicator wasn't within the range of cancer-related genes with high frequency mutations.?6?KEGG signaling pathway analysis showed that the molecular mechanism of GWSMFCP may relate to genes in several signaling pathways including membrane protein and tricarboxylic acid cycle in carbon metabolism,choline phosphorylation metabolic process and PI3K-Akt signaling pathway,and multiple lymphoma transcriptional disorders in choline metabolism.This factor wasn't associated with the p53 signaling pathway which been found commonly in cancer.?7?The results of KOG homologous sequence clustering analysis indicated that GWSMFCP was related to protein post-translational modification and transport,molecular chaperone and signal transduction mechanism,but there were still a large number of functional unclassified genes related to this factor.Conclusions?1?GWSMFCP is a potential molecular marker for the risk of lymph node metastasis of esophageal squamous-cell carcinoma,and has certain clinical application value.?2?GWSMFCP is associated with ubiquitous low-frequency mutational non-driven genes.The functional classification of these genes is mainly focused on signal transduction,protein transport and modification,and gene-unknown functions.