Correlation Between AHNAK And Immune Invasion Of Colorectal Cancer And Its Clinical Significance

Backgroud&Aim:The occurrence,development,invasion,and metastasis of colorectal cancer is a complex process involving multiple stages,multiple steps,and multiple genes,which is one of the most common malignant tumors.AHNAK is a scaffold protein with a size of about 700kDa.desmoyok is its another name.It is reported that AHNAK participates in various tumor pathways,exerts biological effects,and has potential clinical significance as an independent prognostic factor.In this study,TCGA and TIMER databases were employed to analyze the correlation between ANHAK expression in colorectal cancer and immune infiltration,as well as the use of immunohistochemical techniques to detect its expression in clinical specimens in colorectal cancer and liver metastatic cancer tissues,and the role of it in invasion and metastasis in colorectal cancer.Methods:1.Source of specimens:38 tissue specimens of patients with colorectal cancer and liver metastases from colorectal cancer who underwent surgical treatment at the Northern Jiangsu People's Hospital from July 2017 to June 2019.All patients were operated for the first time,and no radiotherapy and chemotherapy were performed before the operation.2.Experimental method:Combined with the analysis of TCGA database to explore the correlation between the expression and prognosis of AHNAK in colorectal cancer,and infiltrating immune cells,including B cells,CD4T cells,CD8T cells,neutrophils,macrophages and immune correlation of dendritic cells,while analyzing the correlation between AHNAK and the expression of immune checkpoint genes in the tumor microenvironment.Finally,immunohistochemical staining was used to study the expression of AHNAK protein in 38 colorectal cancer and liver metastatic cancer specimens and its correlation with clinicopathology.Results:1.The GTEx data set was used to analyze the expression level of the AHNAK gene in various tissues.The database analyzed 308 samples.The results showed that the AHNAK gene was clearly expressed in colorectal tissues.2.Extracted and analyzed colon adenocarcinoma(COAD)samples(158 cases of cancer tissues and 349 normal tissues)and rectal adenocarcinoma(READ)specimens(167 cases of cancer tissues and 10 cases of normal tissues)in the GTEx data set,and analyzed AHNAK gene expression between the two groups showed that compared with normal tissues,AHNAK was highly expressed in colorectal cancer tissues(P<0.001).3.Analysis of the expression of AHNAK in colorectal cancer with B cells,CD4T cells,CD8T cells,neutrophils,macrophages and dendritic cells infiltration by the TIMER database(p<0.05),the results showed that AHNAK and colon cancer gene expression and immune correlation are obvious.4.Furthermore,analyze the relationship between AHNAK gene expression and the expression of immune checkpoint genes,extract these immune checkpoint genes,and calculate the correlation with the expression of AHNAK genes.The results showed that the expression of AHNAK was significantly correlated with common immune checkpoint genes,including CTLA4,CD48,CD28,etc(p<0.05).5.Immunohistochemical staining results showed that the expression rate of ANHAK in colorectal cancer tissues was significantly higher than that in adjacent tissues(p<0.05),and the expression in normal tissues was negative.Immunohistochemical staining results showed that AHNAK protein was mainly expressed in the nucleus,cell membrane and cytoplasm in tumor tissues,and partly expressed in tumor interstitial.6.Additionally,immunohistochemical staining was employed to analyse the correlation between the expression intensity of AHNAK in colorectal cancer tumors and clinicopathology.The results showed that there was no significant correlation between age,sex,tumor location and AHNAK expression intensity(p>0.05).The positive expression rates in well-differentiated,medium-differentiated,and poorly-differentiated cancers were 2/8,9/13,and 15/17,respectively..It indicates that the lower the degree of tumor differentiation,the higher the positive rate of AHNAK expression(p<0.05).7.HE staining was employed to observe the tissue structure of liver metastatic cancer.Compared with normal liver tissue,the glandular structure in metastatic cancer is disordered and the atypical type is more obvious.The expression rate of AHNAK in liver metastatic cancer tissue was analyzed and analyzed by immunohistochemical staining and clinical pathological data.Afterdata analysis,the results showed that AHNAK protein is highly expressed in cancer tissues,but not in normal tissues.8.At the same time,the pathological correlation between the expression of AHNAK and liver metastasis of colorectal cancer was analyzed.The results showed that the positive rate of AHNAK expression in patients with liver metastasis of colorectal cancer was higher than that of patients without colorectal cancer(p<0.05)..There is no obvious correlation between liver metastasis?degree of tumor differentiation?gender and tumor location(p>0.05).Conclusion:High expression of AHNAK in colorectal cancer;AHNAK is highly correlated with immune cell infiltration in tumor tissue;The expression of AHNAK in colorectal cancer is significantly correlated with multiple immune checkpoint genes involved in tumor immunity;AHNAK protein is highly expressed in colon cancer and liver metastatic cancer tissues.