Detection of Marker Associated with CTC in Colorectal Cancer in Mononuclear Cells of Patients with Benign Inflammatory Intestinal Diseases

Abstract

Colorectal carcinoma (CRC) belongs to the most common tumor entities in western countries. Circulating tumor cells (CTC) in blood of CRC patients are a powerful prognostic and predictive biomarker. However, whether CTC-associated markers can also be used for early CRC detection and discrimination from benign diseases is not known. This study investigated the presence of CTC-associated markers CK20, PLS3, LAD1, and DEFA5 in blood of patients with benign inflammatory intestinal disease (IID) and their correlation with malignancy. The detection rate of CK20 and DEFA5 significantly differed between diseased patients and healthy controls. LAD1 and PLS3 were detected in all samples with clear differences in gene expression. DEFA5 expression was higher in CRC and IID patients compared to healthy donors, while CK20 and PLS3 were lower in CRC compared to IID patients or healthy controls. Overall, all CTC-associated markers were detectable in blood of IID patients, but not correlating with inflammation severity. Finally, PLS3 emerged as a suitable marker for differentiation between malignant and non-malignant intestinal diseases or healthy controls, however its suitability for early CRC detection needs to be further validated.

Detection of Circulating and Disseminated Tumor Cells and Their Prognostic Value under the Influence of Neoadjuvant Therapy in Esophageal Cancer Patients

“…To further validate this marker for detection of CTC and DTC in EC patients and with this as an indicator of malignancy, CK20 expression was also assessed in the blood and bone marrow of a control cohort (comprising healthy blood donors and bone marrow donors with non-malignant diseases) showing a positive detection of CK20 in the blood (63%) and bone marrow (42%). Our findings are in line with studies that have also demonstrated CK20 positivity in the blood of healthy donors [ 32 ]. Importantly, it has to be taken into account that all RT-PCR analyses were performed in triplicate with the samples from the EC patients being more often triple or at least double positive for CK20 (but also for DEFA5) than the control cohort.…”

“…In the control group, 57.8% of the blood and 91.6% of the bone marrow samples were positive for DEFA5, which declined to 13% and 45%, respectively, after cut-off calculation. The higher detection rates of DEFA5 in the blood of EC patients compared to healthy controls correspond to the findings for CRC patients published by Born et al, and the findings by Lisitsyn et al, albeit the latter study comprises only a little number of patients and control donors [28,32].…”

“…These findings along with experimental data demonstrating a tumor cell growth inhibitory effect of DEFA5 point to a role of DEFA5 as a tumor suppressor [34,35]. Considering its tumor-suppressive function and its known immunomodulating role of DEFA5, the detection of DEFA5 even in healthy donors is reasonable and has also been reported earlier [32]. Moreover, it may also explain why DEFA5 positivity in blood (EC patients without neoadjuvant therapy) and bone marrow (EC patients with neoadjuvant therapy) is associated with a better outcome of EC patients.…”

“…Since there are still only a few studies [ 19 , 40 ] analyzing the detection of CTC in the blood and DTC in the bone marrow of EC patients, the objective of this study was to analyze the prognostic value of CK20 and DEFA5 mRNA expression levels in blood and bone marrow in a cohort of 216 EC patients. Although the expression of both genes has been described predominantly in the gastrointestinal tract according to the Human Protein Atlas ( (accessed on 24 February 2022)) [ 41 ], and RNA-sequencing data in the Cancer Genome Atlas (TCGA) database indicate that the expression of either gene is not altered in EC compared to the normal tissue [ 42 ], CK20 was chosen as a biomarker for EC cells, because it has already been proven as a good indicator for CTC and DTC in CRC and EC patients with a high prognostic and predictive value [ 19 , 21 , 32 ]. Furthermore, we intended to validate our previous study, in which we analyzed blood and bone marrow samples from a smaller cohort of 77 EC patients [ 19 ].…”

“…One study demonstrated a correlation between elevated plasma levels of α-defensins 1–3 and lymphatic and hepatic metastasis in colorectal cancer (CRC) [ 31 ]. Moreover, we recently demonstrated elevated DEFA5 mRNA expression in blood-derived MNC of CRC patients compared to patients with benign diseases of this organ system and healthy donors [ 32 ]. However, despite some reports on the function of DEFA5 in the regulation of cell growth and apoptosis, its role in tumorigenesis is not yet well understood [ 33 , 34 , 35 ].…”