An Updated Review on Molecular Biomarkers in Diagnosis and Therapy of Colorectal Cancer.

In order to find out the most valuable biomarkers and pathways for diagnosis, therapy and prognosis in colorectal cancer (CRC) we have collected the published CRC biomarkers and established a CRC biomarker database (CBD: http://sysbio.suda.edu.cn/CBD/index.html). In this study, we analysed the single and multiple DNA, RNA and protein biomarkers as well as their positions in cancer related pathways and protein-protein interaction (PPI) networks to describe their potential applications in diagnosis, therapy and prognosis. CRC biomarkers were collected from the CBD. The RNA and protein biomarkers were matched to their corresponding DNAs by the miRDB database and the PubMed Gene database, respectively. The PPI networks were used to investigate the relationships between protein biomarkers and further detect the multiple biomarkers. The Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway enrichment analysis and Gene Ontology (GO) annotation were used to analyse biological functions of the biomarkers. AI classification techniques were utilized to further verify the significances of the multiple biomarkers in diagnosis and prognosis for CRC. We showed that a large number of the DNA, RNA and protein biomarkers were associated with the diagnosis, therapy and prognosis in various degrees in the CRC biomarker networks. The CRC biomarkers were closely related to the CRC initiation and progression. Moreover, the biomarkers played critical roles in cellular proliferation, apoptosis and angiogenesis and they were involved in Ras, p53 and PI3K pathways. There were overlaps among the DNA, RNA and protein biomarkers. AI classification verifications showed that the combined multiple protein biomarkers played important roles to accurate early diagnosis and predict outcome for CRC. There were several single and multiple CRC protein biomarkers which were associated with diagnosis, therapy and prognosis in CRC. Further, AI-assisted analysis revealed that multiple biomarkers had potential applications for diagnosis and prognosis in CRC.

Rates of New or Missed Colorectal Cancers After Colonoscopy and Their Risk Factors: A Population-Based Analysis

Colorectal cancer (CRC) is a disease without evident clinical symptoms in early stages, leading to late diagnosis and disease management. Current diagnostic and prognostic tools require invasive procedures and circulating molecular biomarkers fail to have optimal sensitivity and specificity. Circulating biomarkers with high clinical performance may be valuable for early diagnosis and prognosis of CRC. The purpose of this review was to investigate the application of circulating cell-free DNA (ccfDNA) in CRC diagnosis and prognosis and the analytical methods used in blood samples in articles published between 2005 and 2016. Based on specific inclusion and exclusion criteria, 26 articles were selected. Most studies used ccfDNA quantification as the molecular biomarker. The analytical method was mainly based on the quantitative polymerase chain reaction (qPCR). Biomarkers based on aberrantly methylated genes (n=6) and ccfDNA integrity/fragmentation (n=2) were also used for the CRC diagnosis. The CRC prognosis used the detection of oncogene mutations, such as KRAS and BRAF, in ccfDNA. Significant differences were found in variables among the studies revealing potential bias. ccfDNA quantification as a diagnostic biomarker for CRC has promising results but it lacks clinical specificity since other diseases present a similar increase in ccfDNA content. However, increasing research in the epigenomic field can lead the way to a clinically specific biomarker for the CRC early diagnosis. As for the analytical method, qPCR and derivatives seem to be a perfectly valid technique. The use of ccfDNA quantification in CRC prognosis seems promising. The attempt to use the ccfDNA quantification in clinical practice may reside in the prognosis using a qPCR technique.

We investigated HSP90α as screening biomarker for early colorectal cancer (CRC). Seventy-seven CRC patients and 78 healthy controls were enrolled. Plasma HSP90α was significantly higher in CRC patients than in healthy controls (p<0.05). levels were higher in late (stages III and IV) CRC than in early (stages I and II) CRC (p=0.022). HSP90α conferred an advantage in the diagnosis of early CRC. Combination of HSP90α and carcinoembryonic antigen improved the diagnostic sensitivity (84.4%) and specificity (89.5%) for CRC (area under the curve: 0.968); for early CRC, the sensitivity was 82.5% and specificity was 89.5% (area under the curve: 0.955). HSP90 is a potential biomarker for the diagnosis of early CRC.

Colorectal cancer is one of the common malignant tumors nowadays, with the highest mortality rate among cancers, and most patients have metastasis when diagnosed. Exosomes contain bioactive molecules such as RNA, which can participate in signal transduction between cells. tRF is a new type of non-coding small RNA, which plays an important role in the progression of cancer. The purpose of this study is to explore the value of exosomal tRF as a biological diagnostic marker for colorectal cancer. Through the use of transmission electron microscopy (TEM), qNano, and western blot, exosomes obtained from the serum of both healthy people and patients with colorectal cancer were characterized; the expression of tRNA-Gly-5-0007 was validated by real-time quantitative PCR (qRT-PCR) in the exosome of 172 colorectal cancer patients and 164 healthy people. The diagnostic efficacy of tsRNA-Gly-5-0007 as a diagnostic marker for colorectal cancer was evaluated by receiver operating characteristic curve (ROC) analysis. We further conducted a preliminary verification of how tsRNA-Gly-5-0007 participates in the malignant phenotype of tumors through transwell experiments and cell adhesion experiments. The TCGA database shows that tsRNA-Gly-5-0007 is an RNA fragment derived from tsRNA-Gly-CCC-2-1. The expression of tsRNA-Gly-CCC-2-1 is down-regulated in colorectal cancer tissues and may be involved in Wnt, mTOR and other signaling pathways. tsRNA-Gly-5-0007 can inhibit the adhesion and motor capacity of colorectal cancer cells. Compared with healthy people, the expression of exosomal tsRNA-Gly-5-0007 was significantly down-regulated in patients with colorectal cancer, especially in patients with early colorectal cancer. The diagnostic efficacy of exosomal tsRNA-Gly-5-0007 for the diagnosis of colorectal cancer was 0.7812, and the diagnostic efficacy for the diagnosis of early colorectal cancer was 0.7726. tsRNA-Gly-5-0007 may affect the adhesion and motor capacity of colorectal cancer cells through Wnt signaling pathway, and then participate in the malignant process of colorectal cancer cells. Moreover, the expression of exosomal tsRNA-Gly-5-0007 is down-regulated in colorectal cancer patients and can be used as a promising biomarker for the biological diagnosis of colorectal cancer.

Introduction Recent statistics have reported colorectal cancer (CRC) as the second leading cause of cancer-associated deaths in the world. Early diagnosis of CRC may help to reduce the mortality and associated complications. However, the conventional diagnostic techniques often lead to misdiagnosis, fail to differentiate benign from malignant tissue or diagnose only at an advanced stage. For the treatment of CRC, surgery, chemotherapy, immunotherapy, and radiotherapy have been employed. However, the quality of living of the CRC patients is highly compromised after employing current therapeutic approaches owing to the toxicity issues and relapse. Area covered This review accentuates the molecular mechanisms involved in the pathogenesis, stages of CRC, conventional approaches for diagnosis and therapy of CRC and the issues confronted thereby. It provides an outlook on the advantages of employing nanotechnology-based approaches for prevention, early diagnosis, and treatment of CRC. Expert opinion Employing nanotechnology-based approaches has demonstrated promising outcomes in the prevention, diagnosis, and treatment of CRC. Nanotechnology-based approaches can surmount the major drawbacks of traditional diagnostic and therapeutic approaches. Nanotechnology bestows the advantage of early detection of CRC which helps to undertake instant steps for offering efficient therapy and reducing the mortality rates. For the treatment of CRC, nanocarriers offer the benefit of achieving controlled drug release, improved drug bioavailability, enhanced tumor targetability and reduced adverse effects.

BACKGROUNDEarly diagnosis of colorectal cancer (CRC) is of great significance to improve the survival rate and quality of life of patients, but early diagnosis of CRC requires more sensitive techniques. Peripheral blood UL16-binding protein 2 (ULBP2) and human fibrinogen degradation products (DR-70) are the main indicators for the diagnosis of malignant tumors.AIMTo assess ULBP2 and DR-70 potential for the early diagnosis and prognostic evaluation of CRC to provide a reference.METHODSThis study involved 60 patients with early-stage CRC (CRC group), 50 patients with benign colorectal tumors (benign group), and 50 healthy patients (control group) enrolled at the Affiliated Hospital of Jiangnan University and Jiangsu Province Official Hospital between January, 2020 and January, 2022. ULBP2 and DR-70 levels in the blood were determined and differences among the three groups and early diagnostic values for CRC were determined. Patients with CRC were divided into the good prognosis and poor prognosis groups, and ULBP2 and DR-70 levels in the blood and diagnostic values were compared.RESULTSULBP2 and DR-70 serum levels were significantly higher in the CRC group than in the control and benign groups (P < 0.05); however, no significant differences were observed between the benign and control groups (P > 0.05). Among the 60 patients with CRC followed up for two years, two died (3.33%) and 15 exhibited tumor metastasis, progression, or recurrence (25.00%). ULBP2 and DR-70 serum levels were significantly higher in the poor prognosis group than in the good prognosis group (P < 0.05). A receiver operating characteristic curve was plotted. Area under the curve, sensitivity, and specificity of serum ULBP2 with DR-70 for the early diagnosis of CRC were higher than those of the single serum indices (P < 0.05) in both the good and poor prognosis groups.CONCLUSIONULBP2 and DR-70 serum levels were significantly high in patients with early-stage CRC. They improved the diagnostic rate of early-stage CRC and predicted patient prognosis, thereby showing clinical application potential.

This study aimed to explore critical genes as potential biomarkers for the diagnosis and prognosis of colorectal cancer (CRC) for clinical utility. To identify and screen candidate genes involved in CRC carcinogenesis and disease progression, we downloaded microarray datasets GSE89076, GSE73360, and GSE32323 from the GEO database identified differentially expressed genes (DEGs), and performed a functional enrichment analysis. A protein-protein interaction network was constructed, and correlated module analysis was performed using STRING and Cytoscape. The Kaplan-Meier survival curve shows the survival of the hub genes. The expression of cyclin-dependent kinase (CDK1), cyclin B1 (CCNB1), and PCNA in tissues and changes in tumor grade were analyzed. A total of 329 DEGs were identified, including 264 upregulated and 65 downregulated genes. The functions and pathways of DEGs include the mitotic cell cycle, poly(A) RNA binding replication, ATP binding, DNA replication, ribosome biogenesis in eukaryotes, and RNA transport. Forty-seven Hub genes were identified, and biological process analysis showed that these genes were mainly enriched in cell cycle and DNA replication. Patients with mutations in CDK1, PCNA, and CCNB1 had poorer survival rates. CDK1, PCNA, and CCNB1 were significantly overexpressed in the tumor tissues. The expression of CDK1 and CCNB1 gradually decreased with increasing tumor grade. CDK1, CCNB1, and PCNA can be used as potential markers for the diagnosis and prognosis of CRC. These genes are overexpressed in colon cancer tissues and are associated with low survival rates in CRC patients.

Objective To explore the clinical significance of roundabout guidance receptor 4(ROBO4) and protocadherin 12(PCDH12) in the human serum in the screening,diagnosis,and staging of colorectal cancer(CRC) through proteomics and bioinformatics,thus providing a novel screening method with high specificity,sensitivity,and accuracy for the early screening and diagnosis of CRC in clinical practice. Methods Proteomics and bioinformatics approaches were employed to select the target proteins ROBO4 and PCDH12,and the correlation between ROBO4 and PCDH12 was analyzed.A total of 121 CRC patients(cancer group),112 patients with polyps(polyp group),and 108 healthy volunteers(normal group) who received treatment or physical examination from January 2023 to June 2024 were enrolled in this study.ELISA was employed to determine the expression levels of ROBO4,PCDH12,carcinoembryonic antigen(CEA),and carbohydrate antigen 199(CA199) in the samples of each group.The expression levels of the four proteins in the serum were compared among the three groups of patients mentioned above,and the relationships of the serum levels of ROBO4 and PCDH12 with the pathological characteristics of the CRC patients were analyzed.Furthermore,the receiver operating characteristic(ROC) curves were established to evaluate the diagnostic efficacy of the serum levels of ROBO4 and PCDH12 for CRC patients in early and progressive stages. Results The target proteins ROBO4 and PCDH12 were screened out through proteomics.Bioinformatics analysis showed that ROBO4 and PCDH12 were highly expressed in CRC patients.Further correlation analysis revealed a positive correlation between ROBO4 and PCDH12 in CRC(R=0.870,P<0.001).The ELISA results of clinical samples showed that compared with the normal group and polyp group,the cancer group presented elevated expression levels of ROBO4 and PCDH12(all P<0.001).There was no statistically significant difference in the expression level of ROBO4 or PCDH12 between the normal group and the polyp group(P=0.586,P=0.550).The ROC curves showed that the diagnostic efficacy of ROBO4,PCDH12,and ROBO4+PCDH12 was 0.787,0.757,and 0.812,respectively,all of which were higher than the diagnostic efficacy of CEA and CA199.Further analysis of serum levels of ROBO4 and PCDH12 with the pathological data of CRC showed that the serum level of ROBO4 was correlated with differentiation degree(P=0.013),pathological stage(P=0.002),lymph node metastasis(P=0.001),and distant metastasis(P=0.026).The serum level of PCDH12 was correlated with differentiation degree(P=0.043),pathological stage(P=0.012),and lymph node metastasis(P=0.001).The ROC curves showed that the diagnostic efficacy of ROBO4,PCDH12,and ROBO4+PCDH12 was 0.637,0.758,and 0.787 for early CRC and 0.872,0.757,and 0.882 for progressive CRC,respectively. Conclusions The serum levels of ROBO4 and PCDH12 demonstrate high diagnostic efficacy for CRC patients and are correlated with the pathological features of CRC.The two proteins are expected to be novel serum biomarkers for the diagnosis and screening of CRC.

Race, Ethnicity, and Socioeconomic Status Are Associated With Prolonged Time to Treatment After a Diagnosis of Colorectal Cancer: A Large Population-Based Study

Colorectal cancer (CRC) has been a common malignant tumor in China. Through the great effort in cancer prevention and treatment, the 5-year survival rate of patients with CRC in China has been largely improved. However, insufficient early diagnosis and treatment have seriously impeded the prognosis of Chinese CRC patients. The Early Diagnosis and Treatment Group of the Chinese Medical Association, Oncology Branch organized experts in the area of CRC to write the Expert Consensus on the Early Diagnosis and Treatment of Colorectal Cancer in China and issued it publicly (2020 edition). The 2020 edition of expert consensus has been meaningful to the early diagnosis and standardized treatment of CRC. Based on the prior edition, the Early Diagnosis and Treatment Group of the Chinese Medical Association, Oncology Branch revise the consensus focused on the early screening and diagnosis protocols for colorectal cancer, endoscopic/surgical management of early colorectal tumor, as well as adjuvant treatment and remedial measures and follow-up monitoring protocols. The 2023 edition of the consensus not only synthesized the latest evidence from China and combined it with the specific national condition and clinical practice, but also referred to the international guidelines. This 2023 edition consensus, developed for clinicians working on the screening, early diagnosis and treatment for CRC, would further promote the standardization and advances in the diagnosis and treatment of CRC at the early stage in China, and improve the survival and prognosis of CRC.

Background Colorectal cancer (CRC) is one of the most common causes of cancer-related morbidity and mortality worldwide. The most widely used CRC diagnostic tests either noninvasive or invasive are limited by insufficient performance or patient compliance. Profiling inflammatory mediators with carcinogenic roles could aid in CRC diagnosis. We aimed to use multiplex bead assay in evaluating the utility of serum eotaxin-1, macrophage-inflammatory protein-1β (MIP-1β), granulocyte colony-stimulating factor (G-CSF), vascular endothelial growth factor A (VEGF-A), and Fas ligand (FasL) as potential biomarkers in CRC. Patients and methods The study was conducted on 87 patients. Based on colonoscopy findings, patients were divided into 35 CRC and 52 nonmalignant patients (nine with colon polyp, 24 with colitis, and 19 with normal mucosa). Multiplex assay was used for measuring the studied biomarkers. Results The median values of eotaxin-1, MIP-1β, G-CSF, and VEGF-A were significantly higher in patients with CRC compared with the nonmalignant group. The area under the receiver operating characteristics curve for eotaxin-1, MIP-1β, G-CSF, and VEGF-A was 0.863. The area under the ROC for occult blood in stool was only 0.597. Moreover, significantly higher levels of G-CSF and VEGF-A were found in patients with CRC than the precancerous colon polyp group. Conclusions Serum profiling of eotaxin-1, MIP-1β, G-CSF, and VEGF-A could be used as potential biomarkers in early CRC diagnosis with better discriminatory power than stool occult blood and may increase occult blood performance, thus reducing false-positives rates and unneeded colonoscopy. Multiplexing bead technology represents a promising approach for CRC screening and diagnosis.

BackgroundAs a new epigenetic biomarker, 5‐hydroxymethylcytosine (5hmC) is broadly involved in various diseases including cancers. However, the function and diagnostic performance of 5hmC in colorectal cancer (CRC) remain unclear.ResultsHigh‐throughput sequencing was used to profile 5hmC levels in adjacent normal colon, advanced adenomas, and CRC. The expression and 5hmC levels in zw10 kinetochore protein (ZW10) were significantly increased in the tissues and blood samples for patients with advanced adenoma and CRC, and were much higher in the early stages of CRC (I and II). The receiver operating characteristic analysis had potential diagnostic value for CRC. The area under the curve (AUC) of ZW10 5hmC levels in tissue samples of CRC was 0.901. In blood samples, the AUC was 0.748 for CRC. In addition, the ZW10 5hmC level had much higher diagnostic performance in early stages of CRC (AUC = 0.857) than it did in advanced stages (AUC = 0.594). Compared with FHC cell, ZW10 expression in HT29 cell was significantly increased. The ZW10 knockdown could inhibit cell proliferation and the ZW10 overexpression could promote cell proliferation in HT‐29 cell. Furthermore, ZW10 knockdown inhibited AKT and mTOR phosphorylation, and ZW10 overexpression promoted AKT and mTOR phosphorylation.ConclusionsThe ZW10 5hmC level may serve as an effective epigenetic biomarker for minimally invasive screening and diagnosis of CRC, and it has higher diagnostic performance in early stages of CRC than it does in advanced stages. In addition, ZW10 could regulate CRC progression through the AKT‐mTOR signaling.

Early diagnosis and treatment of colorectal cancer (CRC) are important for improving patients' survival. Metadherin is an oncogene that plays a pivotal role in carcinogenesis and can be suggested as a cancer biomarker. This study aimed to elucidate the efficacy of serum Metadherin mRNA expression as a potential non-invasive biomarker for early diagnosis of CRC in relation to other screening markers as carcinoembryonic antigen (CEA), carbohydrate antigen 19.9 (CA19.9) and Fecal occult blood (FOB) and also to assess its relationship with the tumor stage and survival rate. A convenience series of 86 CRC cases (group I) were recruited with 78 subjects as controls (group II). Serum Metadherin mRNA expression level was determined using reverse transcription polymerase chain reaction (RT-PCR). Serum Metadherin mRNA expression level was significantly elevated in CRC cases when compared with controls (P < 0.001). For CRC diagnosis; Receiver operator characteristic (ROC) analyses revealed that the diagnostic accuracy of serum Metadherin mRNA (AUC = 0.976) was significantly higher than other routine CRC screening markers as CEA, CA19.9 and FOB. The combined accuracy of these markers (AUC = 0.741) was increased when used with serum Metadherin mRNA (AUC = 0.820). High serum Metadherin mRNA expression was associated with poorly differentiated histological grade, advanced tumor stage and lower survival rate. AUC of Metadherin was 0.820 for differentiating advanced versus early tumor stages. Serum Metadherin mRNA expression is a useful non-invasive biomarker for CRC. It can be used for screening and early diagnosis of CRC and can increase the efficacy of other routine CRC screening markers when it is estimated in CRC patients with them. It is also associated with advanced tumor stage and a lower survival rate.

Objective To study the analysis of SEPT9 gene methylation in diagnosis and follow-up of colorectal cancer. Methods The clinical data of 783 cases of colorectal cancer screening in our hospital from October 2014 to December 2015 were analyzed. There were 72 patients diagnosed with colorectal cancer, 711 cases were excluded colorectal cancer. In there, all patients were performed by peripheral blood SEPT9 methylation detection and pathological examination. Pathological diagnosis was used as gold standard, with the ROC curve, the diagnostic value of SEPT9 positive for colorectal cancer was evaluated. After 1 years of follow-up, the SEPT9 gene methylation and the survival of the patients were observed. Statistical analysis were performed by using SPSS 23.0 software, the classification data such as sex, age, location of lesion, TNM stage and et al were presented as n(%), and examined by using Chi square test. A P<0.05 was considered as significant difference. Results The level of SEPT9 gene methylation of colorectal cancer patients was significant higher than non-colorectal cancer patients. The positive rate of SEPT9 gene methylation of colorectal cancer patients in stage Ⅰ, stage Ⅱ, stage Ⅲ and Ⅳ was 47.4%, 69.2%, 88.89%, 88.9% respectivly. ROC curve analysis showed that the SEPT9 was higher than that of CA125 and CA199 curve (P<0.05), the sensitivity and specificity of the diagnosis of colorectal cancer was 86.5% and 70.8%. After 1 year of follow-up, 88.2% of positive SEPT9 gene methylation patients turned to negative after colorectal cancer surgery, compared with this patients, the prognosis of patients whose SEPT9 methylation had not turned to negative had a poor prognosis. Conclusion SEPT9 gene methylation could be used as an important test indicator for the risk and prognosis of colorectal cancer. Key words: Colorectal neoplasms; ROC curve; SEPT9 gene