| Objective:Esophageal carcinoma(ECA)is one of the most common malignant tumors of the digestive system,and the main pathological type is esophageal squamous cell carcinoma(ESCC).Although there are various treatments for esophageal cancer,including surgery,radiotherapy,chemotherapy and immunotherapy,the five-year survival rate is still low.In recent years,it has been found that targeted therapy has good efficacy and low toxicity,so exploring meaningful targets is expected to benefit more patients with esophageal cancer.Nectin cell adhesion molecule 1(Nectin-1)is one of the four members of the immunoglobulin-like cell adhesion molecule family,which promotes the formation of intercellular adhesion and regulates a series of cellular activities.In recent years,abnormal expression of Nectin-1 has been reported in gastric cancer,colorectal cancer,pancreatic ductal adenocarcinoma,etc,but the relationship between Nectin-1 and esophageal cancer has not been fully clarified.This study aims to explore the role of Nectin-1 expression in ESCC and its possible mechanism.It is hoped to provide new markers for the diagnosis of ESCC and bring new ideas for the development of new targeted drugs.Methods:(1)six cancer and paracancer tissue samples from patients undergoing surgery for esophageal squamous cell carcinoma were selected for mRNA sequencing screening to obtain differentially expressed genes and preliminarily screen out key molecules.(2)Using biological information technology to analyze the expression of Nectin-1 and its clinical significance:①Using TIMER database to analyze the expression of Nectin-1 gene in malignant tumors;②Kaplan-Meier Plotter database was used to analyze the relationship between Nectin-1 expression and prognosis of esophageal cancer.(3)TIMER database was used to analyze the correlation between Nectin-1 and special immune cells,programmed death receptor 1(PD-1)and programmed death ligand 1(PD-L1)in esophageal cancer.(3)Biological function of Nectin-1 in ESCC was investigated by cell assay:①Esophageal squamous cell carcinoma cells with high Nectin-1 expression were screened by qPCR and Western blot experiment;②After the interference of Nectin-1 expression by lentivirus,CCK-8 cell proliferation experiment,Cell scratch experiment,Transwell migration experiment,Transwell invasion experiment and Flow cytometry cycle experiment were performed.(4)Using Western blot experiment to explore the mechanism of Nectin-1 in ESCC:Western blot experiment was performed to detect the relative expression of major proteins of PI3K/AKT,STAT3 and MAPK(ERK/P38/JNK)signaling pathways in the Nectin-1 knockdown group group and the control group.Results:(1)The key gene Nectin-1 was obtained by mRNA sequencing combined with TIMER database screening.In the kaplan-Meier Plotter online database,it was found that high expression of Nectin-1 in ESCC had worse overall survival(OS)and recurrence free survival(RFS),although there was no statistical significance(P>0.05),the curve separation was obvious.TIMER database showed that Nectin-1 was associated with special immune cells and immune checkpoints in esophageal cancer,and Nectin-1 may play a certain role in immunotherapy.(2)Interference with Nectin-1 expression significantly inhibited the proliferation,migration and invasion of ESCC,and blocked esophageal squamous carcinoma cells in G1 phase,suggesting that Nectin-1 may interfere with the proliferation,migration and invasion of esophageal squamous carcinoma cells at the cellular level,and affect the cycle of ESCC.(3)The expression of phosphorylated proteins of PI3K/AKT,STAT3,ERK/P38/JNK decreased after Nectin-1 expression was disrupted.Conclusion:Nectin-1 is highly expressed in ESCC,which may regulate its proliferation,migration,invasion and cell cycle through PI3K/AKT,SATA3 and MAPK signaling pathways. |
