Effect Of ILK On Malignant Phenotype And Function Of Esophageal Squamous Cell Carcinoma

Objective: The purpose of this study was to investigate the effect of integrin-linked kinase(ILK)gene on the proliferation,apoptosis,invasion and migration of esophageal cancer cells,and to study its function in esophageal cancer,so as to lay a theoretical foundation and clinical reference for accurate diagnosis and treatment of esophageal cancer.Methods: 1)meta analysis was used to evaluate the expression of ILK in digestive system tumors and its correlation with clinical phenotype and prognosis of digestive system tumors.2)Human esophageal squamous cell carcinoma lines with high expression of ILK were screened to establish ILK lentivirus interference sequence.The ILK lentivirus interference sequence was transfected into esophageal squamous cell carcinoma cell lines TE-1 and KYSE-150,The expression of ILK after transfection was detected by q RT-PCR.At the cellular level,MTT proliferation,clone detection,flow cytometry apoptosis detection,Transwell chamber invasion and scratch healing were used to study the effects of ILK gene deletion on the proliferation,apoptosis,invasion and migration of esophageal squamous cell carcinoma cells.3)establish ILK lentivirus interference vector and transfect KYSE-150 cells.Sh ILK and control cell lines were inoculated subcutaneously in nude mice to establish the subcutaneous transplanted tumor model of KYSE-150 cells in nude mice.The tumor growth was observed and the size and volume of the tumor were measured.To study the effect of ILK gene deletion on the growth of esophageal squamous cell carcinoma at animal level.The differentially expressed genes of esophageal cancer were screened based on RNA-seq.The possible pathogenesis of esophageal cancer was predicted through GO function enrichment and KEGG pathway analysis,and the key regulatory genes and related signal pathways that may be involved in the occurrence and development of esophageal cancer were screened.Results: Part1: the results of meta analysis showed that ILK was highly expressed in digestive system tumors.The expression of ILK was correlated with tumor differentiation,TNM stage,lymph node metastasis,tumor invasion and survival prognosis,but not with age and sex.Part 2: at the level of ESCC cells in vitro,silencing ILK gene could significantly inhibit the proliferation and colony formation of TE-1 and KYSE-150 cells,and promote the apoptosis of TE-1 and KYSE-150 cells.After the deletion of ILK gene,the invasion and migration ability of TE-1 and KYSE-150 cells decreased significantly.Part3:the results of the animal model of transplanted tumor in nude mice showed that the tumor in the ILK interference group grew slowly,and the weight and volume of the tumor were smaller than those in the control group.The subtraction of ILK gene affected the ability of tumor formation in nude mice.Based on RNA-seq technique,5540 differentially expressed genes in esophageal cancer were found after ILK gene interference,including 2839up-regulated genes and 2601 down-regulated genes.The up-regulated genes are SOD2,IRF6,PER1,PHLDB2,TNFAIP3,etc.while the down-regulated genes are RPL21,HYOU1,WARS,SUPT16 H,DHCR24 and so on.It may be involved in TNF signal pathway,AMPK signal pathway,Fox O signal pathway,p53 signal pathway,NF-k B signal pathway and so on.Conclusion: The high expression of ILK in digestive system tumors was correlated with tumor differentiation,TNM stage,lymph node metastasis,tumor invasion and survival prognosis,but not with age and sex.ILK gene deletion inhibits the proliferation,clone formation,invasion and metastasis of ESCC cells,and promotes apoptosis.In vivo tumorigenesis studies have confirmed that the deletion of ILK gene will hinder the growth of KYSE-150 transplanted tumor.The differentially expressed genes in esophageal cancer were screened based on RNA-seq technique,and the differentially expressed genes and possible related signal transduction pathways were predicted by GO functional enrichment and KEGG pathway analysis.