Screening Of Potential DNA Methylation Markers In Early-stage Colorectal Cancer

BACKGRANDColorectal Cancer(CRC)is one of the most common cancers with high incidence and mortality.Clinically,early CRC patients have no specific clinical manifestations,resulting in low rate of early diagnosis and treatment;when the disease progresses to the middle and late stage,the prognosis is poor and the fatality rate is high.The existing structural screening methods are invasive examination,the missed diagnosis rate of adenoma is high,and most of the patients with symptoms belong to the late stage,which is not conducive to the promotion of early screening and early diagnosis.The sensitivity and specificity of fecal occult blood test,fecal DNA and plasma mSEPT9 detection for early lesions are not high.There is an urgent need to find new screening and diagnostic markers for CRC.OBJECTIVEIn the preliminary work with our partner Anchor DX,we selected seven DNA methylation biomarkers ZNF132,KCNQ5,FBN1,C9orf50,NEUROD1,ST8SIA4 and FAM72C.We compared the methylation levels of inflammatory bowel disease,benign precancerous lesions,high-grade intraepithelial neoplasia and colorectal cancer to determine how epigenetic changes may promote cancer formation.The further purpose of this study was to detect 7 genes in different subtypes and non-cancerous tissues of colorectal cancer in order to distinguish between non-cancerous tissues and colorectal lesions,as well as methylated biomarkers with good specificity and sensitivity in colorectal benign precancerous lesions,and to establish a model for predicting the diagnosis of colorectal precancerous lesions and colorectal cancer.MATERIAS AND METHODS88 colorectal tissue samples were collected,including inflammatory Bowel Disease,benign precancerous lesions,high-grade intraepithelial neoplasia and colorectal cancer.Seven methylated markers selected by previous studies were verified by MethyLight in colorectal lesion tissue samples.It is used for the detection and screening of precancerous lesions and early colorectal cancer,and the classification model is further used to evaluate the clinical performance of these biomarkers.RESULTSZNF132,KCNQ5,FBN1,C9orf50,FAM72C and ST8SIA4 markers are relatively good for distinguishing Non-CRC from CRC and Early CRC.The area under the curve(AUC)of the six methylated markers to identify CRC is 0.87-0.95,the sensitivity is 72.55%-94.12%,and the specificity is 86.49-97.3%.The AUC for identifying Early CRC is 0.89-0.95,the sensitivity is 77.78%-97.22%,and the specificity is 83.78-100%.and the specificity of ST8SIA4 for identifying early CRC is 100%.The combination classifier of ZNF132 and FBN1 genes has the best performance and stability for recognizing CRC.The AUC of the classifier is 0.94,the sensitivity is 89.6%,and the specificity is 93.3%.The double marker(NEUROD1 and FAM72C)model was related to CPL.The AUC of CPL model for distinguishing IBD group from CPL group was 0.87,the sensitivity was 83.2%,and the specificity was 77.4%.When the prevalence rate of CPL was 25%,the negative predictive value of CPL model was 93.3%.CONCLUTIONS1.The methylation markers ZNF132,FBN1,NEUROD1,FAM72C,KCNQ5,C9orf50 and ST8SIA4 used to identify CPL,Early CRC and CRC lesions were preliminarily identified at the tissue level.2.The combination of ZNF132 and FBN1 markers has the best performance for identifying CRC.The AUC of the classifier is 0.94,the sensitivity is 89.6%,and the specificity is 93.3%.It has the potential for early screening and early diagnosis of colorectal cancer.3.The AUC of NEUROD1 and FAM72C in distinguishing IBD from CPL is 0.87,the sensitivity and specificity are 83.2%and 77.4%.When the prevalence rate of CPL is 25%,NPV is estimated to be 93.3%.It has the potential to become a marker for screening early colorectal lesions in order to reduce the use of colonoscopy.