Circulating Tumor Cells Adhesion: Application in Biosensors.

The early and non-invasive diagnosis of tumor diseases has been widely investigated by the scientific community focusing on the development of sensors/biomarkers that act as a way of recognizing the adhesion of circulating tumor cells (CTCs). As a challenge in this area, strategies for CTCs capture and enrichment currently require improvements in the sensors/biomarker's selectivity. This can be achieved by understanding the biological recognition factors for different cancer cell lines and also by understanding the interaction between surface parameters and the affinity between macromolecules and the cell surface. To overcome some of these concerns, electrochemical sensors have been used as precise, fast-response, and low-cost transduction platforms for application in cytosensors. Additionally, distinct materials, geometries, and technologies have been investigated to improve the sensitivity and specificity properties of the support electrode that will transform biochemical events into electrical signals. This review identifies novel approaches regarding the application of different specific biomarkers (CD44, Integrins, and EpCAm) for capturing CTCs. These biomarkers can be applied in electrochemical biosensors as a cytodetection strategy for diagnosis of cancerous diseases.

A Previously Unrecognized Molecular Landscape of Lynch Syndrome in the Mexican Population.

Lynch syndrome (LS) is the main hereditary colorectal cancer syndrome. There have been few reports regarding the clinical and molecular characteristics of LS patients in Latin America; this is particularly true in the Mexican population, where no information is available. The present study aims to describe the clinical and molecular spectrum of variants in a cohort of patients diagnosed with LS in Mexico. We present a retrospective analysis of 412 patients with suspected LS, whose main site of cancer diagnosis was the colon (58.25%), followed by the endometrium (18.93%). Next-generation sequencing analysis, with an extensive multigene panel, showed that 27.1% (112/414) had a variant in one of the genes of the mismatch repair pathway (MMR); 30.4% (126/414) had a variant in non-MMR genes such as CHEK2, APC, MUTYH, BRCA1, and BRCA2; and 42.5% (176/414) had no genetic variants. Most of the variants were found in MLH1. Pathogenic variants (PVs) in MMR genes were identified in 65.7% (96/146) of the total PVs, and 34.24% (45/146) were in non-MMR genes. Molecular and clinical characterization of patients with LS in specific populations allowed personalized follow-up, with the option for targeted treatment with immune checkpoint inhibitors and the development of public health policies. Moreover, such characterization allows for family cascade testing and consequent prevention strategies.

Evaluation of cancer stem cells markers expression in HCC trough real-time polymerase chain reaction.

Quantitative real-time polymerase chain reaction (qRT-PCR) flexibility, robustness and reproducibility have rapidly extended the scope of the method. Cancer stem cells are gaining increasing importance since their role in cancer initiation, treatment resistance and recurrence give rise to a wide range of potential diagnostic and therapeutic applications. The expression of several characteristic markers is proven a reliable method to assess stem-like-phenotype of cancer cells. Here, we provided a thorough protocol for the study of cancer stem cells in hepatocellular carcinoma mouse models and cell cultures using qRT-PCR.

EpCAM expression in canine malignant mammary tumors - immunohistochemical study

Background: Epithelial cell adhesion molecule (EpCAM) is a glycoprotein responsible for multiple cellular functions that includes cell-to-cell adhesion and signal transduction. Studies in human breast cancer demonstrated that its overexpression has been linked to a more aggressive histological phenotype. However, in canine mammary tumors, its use is still scarce and its function, remains unknown. The main goal of the present study was to characterize the expression of EpCAM in canine mammary malignant tumors and associate its expression with clinicopathological features. Materials, Methods & Results: Forty-eight canine mammary samples were analyzed in the present study and went through the Hematoxylin-Eosin (HE) routine technique for histopathological and immunohistochemical analysis. EpCAM expression was evaluated considering the total immunolabelling (TI) which consists of the addition of 2 parameters: proportion score (PS) and intensity score (IS). Thirteen (27.1%) cases showed weak expression, 9 (18.7%) were characterized by a moderate expression and 27 (54.2%) were classified as intense. EpCAM overexpression was described in 36 (72.9%) primary tumors and in 5 lymph node metastases (71.4%). Immunoreactivity was, mainly, characterized by a staining in the lateral cell membrane of luminal epithelial cells, both in primary tumors and lymph node metastases. The statistical analysis was performed using the SPSS Statistics, version 24. Clinicopathological features, such as histological grade (P = 0.027), intravascular emboli (P = 0.004) and lymph node metastases (P = 0.016) demonstrated to be associated with a more aggressive histological type. Intravascular emboli and lymph node metastases were also linked to a higher histological grade. Discussion: EpCAM overexpression has been described in the literature both in canine mammary tumors and breast cancer. While the results in breast cancer could be ambiguous, depending on the tumor subtype, in the present study our findings demonstrated that overexpression was present in more than 50% of the total cases evaluated. In addition, immunoreactivity was also in accordance with the literature that describes membranous staining as the most prevalent pattern type. It was not possible to correlate overexpression with histological type and histological grade. However, those clinicopathological features are essential to evaluate tumor prognosis. For instance, our results demonstrated that histological type was statistically significant with histological grade (P = 0.027), intravascular emboli (P = 0.004) and lymph node metastases (P = 0.016). Additionally, the Nottingham system was statistically significant with intravascular emboli (P = 0.033) and lymph node metastases (P = 0.006). These findings have also been described in similar studies using canine mammary tissues in bitches. When it comes to pattern of expression in primary tumors and lymph nodes metastases, our findings described that neoplastic cell presented the same phenotype in these 2 different locations. In addition, loss of expression was also described in cluster cells within the subcapsular area which reflects the idea that the expression of EpCAM molecule changes according to time, which could be translated into the multistep process of metastases. Finally, it demonstrates that the development of neoplasia is not something static, but rather dynamic.

EpCAM Expression in Lymph Node and Bone Metastases of Prostate Carcinoma: A Pilot Study.

There is an urgent need for new imaging modalities in prostate carcinoma staging. A non-invasive modality that can assess lymph node and bone metastases simultaneously is preferred. Epithelial cell adhesion molecule (EpCAM) is a membranous protein of interest as an imaging target since it is overexpressed in prostatic carcinoma compared with benign prostate epithelium and compared with stroma. However, EpCAM expression in lymph node metastases is sparsely available in the literature and EpCAM expression in bone metastases is yet unknown. The current study evaluates the expression of EpCAM in prostate carcinoma lymph nodes, in matched normal lymph nodes, in prostate carcinoma bone metastases, and in normal bone by immunohistochemistry. EpCAM was expressed in 100% of lymph node metastases (21 out of 21), in 0% of normal lymph nodes (0 out of 21), in 95% of bone metastases (19 out of 20), and in 0% of normal bone (0 out of 14). Based on these results, EpCAM may be a feasible imaging target in prostate carcinoma lymph node and bone metastases. Prospective clinical trials are needed to confirm current results. Preoperative visualization of prostate carcinoma metastases will improve disease staging and will prevent unnecessary invasive surgery.

Enteropatia com formação de tufos epiteliais e mutação do gene EpCAM: relato de caso / Tufting enteropathy with EpCAM mutation: case report

Tufting enteropathy (TE), also known as intestinal epithelial dysplasia (IED), is a rare congenital enteropathy related to an earlyonset of severe intractable diarrhea due to specific abnormalities of the intestinal epithelium and mutations of the EpCAM gene. TE is characterized by clinical and histological heterogeneity, such as with low or without mononuclear cell infiltration of the lamina propria, and abnormalities of basement membrane. TE can be associated with malformations, other epithelial diseases, or to abnormal enterocytes development and/or differentiation. The authors report a case of a Brazilian child with TE associated with c.556-14A>G mutation in the EpCAM gene (NM_002354.2)...

Enteropatia com formação de tufos epiteliais (ETE), também conhecida como displasia epitelial intestinal (DEI), é uma rara enteropatia congênita relacionada com um início precoce de diarreia intratável grave devido a anormalidades específicas do epitélio intestinal e mutações do gene EpCAM. ETE caracteriza-se por uma heterogeneidade clínica e histológica, como ausência ou leve infiltrado de células mononucleares na lâmina própria e anormalidades de membrana basal. Pode ser associada a malformações, outras doenças epiteliais ou anormalidades no desenvolvimento/na diferenciação dos enterócitos. Os autores relatam um caso de ETE, em uma criança brasileira, associada à mutação c.556-14A> g do gene EPCAM (NM_002354.2)...