Keratin 17 Activates AKT Signaling And Induces EMT To Promote Proliferation And Metastasis Of Esophageal Squamous Cell Carcinoma

Objective: Esophageal squamous cell carcinoma(ESCC)is an aggressive malignancy and a leading cause of cancer-related death worldwide.Lack of e ective early diagnosis strategies and ensuing complications from tumor metastasis account for the majority of ESCC death.Thus,there is an urgent need to identify more sensitive and specific molecular markers involved in the progression of ESCC and to reveal the molecular mechanisms of the occurrence and development of ESCC to improve the early diagnosis and prognosis of ESCC.Methods: 1.Four pairs of ESCC tissues were used for mRNA sequencing to determine di erentially expressed genes(DEGs),and the bioinformatics analysis was performed on DEGs.2.RT-q PCR and Western Blotting were used to detect the expression of KRT17 in 12 pairs of ESCC tissues.And 64 pairs of ESCC were detected by IHC combined with clinical data analysis,we revealed the effect of KRT17 expression on the prognosis of ESCC patients.3.KRT17 overexpression and knockout stable transfected ESCC cell lines were constructed using lentiviral overexpression technology and Crisper-Cas9 technology,and KRT17 upregulated and knockout ESCC cells were verified by Western Blotting.4.CCK8,plate clone formation experiment and subcutaneous tumor formation experiment in BALB/c nude mice were used to detect the effect of KRT17 on the ESCC cell proliferation;Wound-healing,trans-well assay,and tail vein injection experiment in BALB/c nude mice were to detect the influence of KRT17 on ESCC cell migration.5.RT-q PCR and Western Blotting were used to detect the AKT signaling pathway-related proteins(pan-AKT,473ser-AKT)and EMT related proteins(E-cadherin,ZO-1,Vimentin,N-cadherin,snail);Finally,the relationship between AKT pathway activation and EMT was analyzed by sequencing data of 81 cases of ESCC from TCGA database.Results: 1.Through mRNA high-throughput sequencing,it was found that 347 genes were up-regulated and 255 genes were down-regulated in ESCC.Combined with bioinformatics analysis(PPI,GO,KEGG,IPA),protein-protein independent interactions were found in the KRT family.2.High expression of KRT17 and low expression of KRT13 were found in ESCC.Combined with clinical data,high expression of KRT17 was closely related to tumor stage and poor prognosis of ESCC patients.3.Function experiments found that compared with the negative control group,the proliferation and migration capabilities of ESCC cells were significantly improved after KRT17 overexpression;After KRT17 knockout,the proliferation and migration of ESCC cells were inhibited.4.High expression of KRT17 in ESCC promotes the activation of the AKT signaling pathway and is positively correlated with the occurrence of EMT,and 81 cases of TCGA database found that activation of the AKT pathway in ESCC is closely related to EMT.Conclusion: 1.KRT17 is highly expressed in ESCC and is closely related to the tumor stage and poor prognosis of ESCC patients.2.KRT17 promotes the proliferation and metastasis of ESCC by activating the AKT signaling pathway and inducing EMT.KRT17 is closely related to the malignant progression of patients with ESCC and may serve as a new biological target for the early diagnosis and treatment of ESCC.