CD133/CD166/Ki-67 Triple Immunouorescence Assessment for Putative Cancer Stem Cells in Colon Carcinoma*

Background & Aims: Colorectal cancer represents the third most common malignancy and the fourth most common cause of cancer death worldwide. The existence of drug-resistant colon cancer stem cells is thought to be one of the most important reasons behind treatment failure in colon cancer, their existence putatively leading to metastasis and recurrences. The aim of our study was to investigate the immunoexpression patterns of CD133 and CD166 in colon carcinoma, both individually and in combination, assessing their significance as prognostic markers.Methods. A total of 45 retrospective colon adenocarcinoma cases were investigated by enzymatic and multiple fluorescence immunohistochemistry for their CD133 and CD166 expression and colocalization.Results. Both CD133 and CD166 were expressed to different extents in all cancer specimens, with apredominant cytoplasmic pattern for CD133 and a more obvious membranous-like pattern for CD166.Overall, when comparing their reactivity for the tumoral tissue, CD166 expression areas seemed to be smaller than those of CD133. However, there was a direct correlation between CD133 and CD166 expression levels throughout the entire spectrum of lesions, with higher values for dysplastic lesions. Colocalization of CD133/ CD166 was obvious at the level of cells membranes, with higher coeficients in high grade dysplasia, followed by well and moderate differentiated tumours.Conclusions. CD133/CD166 colocalization is an early event occurring in colon tumorigenesis, with thehighest coeficients recorded for patients with high grade dysplasia, followed by well differentiated tumours. Thus, we consider that the coexpression of these two markers could be useful for further prognostic andtherapeutically stratification of patients with colon cancer.Abbreviations: AJCC - American Joint Committee on Cancer; CCD - charge-coupled device camera sensor; CD133 - prominin-1 (PROM1); CD166 - Activated Leukocyte Cell Adhesion Molecule (ALCAM); CRC - colorectal cancer; CSC - cancer stem cells; DAB - 3,3'-diaminobenzidine chromogen; DAPI - 4',6-diamidino- 2-phenylindole; HE - Hematoxylin and eosin staining; HGD - high grade dysplasia; HRP - horseradish peroxidase; LGD - low grade dysplasia; SDS - sodium dodecyl sulfate*Part of this work has been accepted as a poster presentation at the Digestive Disease Week (DDW) meeting, Chicago, IL, USA May 3-6, 2014

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Cytometric Profiling of CD133+ Cells in Human Colon Carcinoma Cell Lines Identifies a Common core Phenotype and Cell Type-specific Mosaics

The International Journal of Biological Markers ◽

10.5301/jbm.5000020 ◽

2013 ◽

Vol 28 (3) ◽

pp. 267-273 ◽

Cited By ~ 11

Author(s):

Marica Gemei ◽

Rosa Di Noto ◽

Peppino Mirabelli ◽

Luigi Del Vecchio

Keyword(s):

Colorectal Cancer ◽

Stem Cells ◽

Cancer Stem Cells ◽

Colon Carcinoma ◽

Cell Lines ◽

Carcinoma Cell ◽

Cell Type ◽

Carcinoma Cell Lines ◽

Cell Type Specific

In colorectal cancer, CD133+ cells from fresh biopsies proved to be more tumorigenic than their CD133– counterparts. Nevertheless, the function of CD133 protein in tumorigenic cells seems only marginal. Moreover, CD133 expression alone is insufficient to isolate true cancer stem cells, since only 1 out of 262 CD133+ cells actually displays stem-cell capacity. Thus, new markers for colorectal cancer stem cells are needed. Here, we show the extensive characterization of CD133+ cells in 5 different colon carcinoma continuous cell lines (HT29, HCT116, Caco2, GEO and LS174T), each representing a different maturation level of colorectal cancer cells. Markers associated with stemness, tumorigenesis and metastatic potential were selected. We identified 6 molecules consistently present on CD133+ cells: CD9, CD29, CD49b, CD59, CD151, and CD326. By contrast, CD24, CD26, CD54, CD66c, CD81, CD90, CD99, CD112, CD164, CD166, and CD200 showed a discontinuous behavior, which led us to identify cell type-specific surface antigen mosaics. Finally, some antigens, e.g. CD227, indicated the possibility of classifying the CD133+ cells into 2 subsets likely exhibiting specific features. This study reports, for the first time, an extended characterization of the CD133+ cells in colon carcinoma cell lines and provides a “dictionary” of antigens to be used in colorectal cancer research.

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Increased risk of high-grade dysplasia and colorectal cancer in inflammatory bowel disease patients with recurrent low-grade dysplasia

Gastrointestinal Endoscopy ◽

10.1016/j.gie.2019.12.041 ◽

2020 ◽

Vol 91 (6) ◽

pp. 1334-1342.e1 ◽

Cited By ~ 2

Author(s):

Michiel E. de Jong ◽

Heleen Kanne ◽

Loes H.C. Nissen ◽

Joost P.H. Drenth ◽

Lauranne A.A. P. Derikx ◽

...

Keyword(s):

Colorectal Cancer ◽

Inflammatory Bowel Disease ◽

Bowel Disease ◽

High Grade Dysplasia ◽

Low Grade ◽

High Grade ◽

Increased Risk ◽

Inflammatory Bowel ◽

Low Grade Dysplasia

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P083. Low-grade Dysplasia in Ulcerative Colitis: Risk Factors associated with Progression to High-grade Dysplasia or Colorectal Cancer

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jju027.201 ◽

2015 ◽

Vol 9 (suppl 1) ◽

pp. S119-S120

Keyword(s):

Colorectal Cancer ◽

Risk Factors ◽

Ulcerative Colitis ◽

High Grade Dysplasia ◽

Low Grade ◽

High Grade ◽

Factors Associated ◽

Low Grade Dysplasia

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Screening for gastrointestinal disease

10.1093/med/9780199568741.003.0354 ◽

2018 ◽

Author(s):

Satish Keshav ◽

Alexandra Kent

Keyword(s):

Colorectal Cancer ◽

Early Detection ◽

Advanced Disease ◽

Gastrointestinal Disease ◽

Hepatocellular Cancer ◽

High Grade Dysplasia ◽

Low Grade ◽

High Grade ◽

The Impact

This chapter discusses screening for gastrointestinal disease, including Barrett’s oesophagus (BO), colorectal cancer, and hepatocellular cancer (HCC). In patients with BO, approximately 5% will develop dysplasia, and 10%–50% of the low-grade dysplasias will progress to high-grade dysplasia or adenocarcinoma within 2–5 years. Thus, screening for BO has been developed to reduce the development of adenocarcinoma via the early detection of high-grade dysplasia or cancer in situ. The main aim of colorectal cancer screening is the early detection of polyps and cancers, at a time when treatment is likely to be more effective. Similarly, early detection of HCC is advantageous, as the prognosis in advanced disease is very poor. This chapter describes the current processes of screening for these diseases, and the impact of this screening, as well as screening for gastrointestinal cancer in specific groups.

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Getting a Low Grade for Missing High-Grade Dysplasia and Colorectal Cancer in IBD

Digestive Diseases and Sciences ◽

10.1007/s10620-017-4795-4 ◽

2017 ◽

Vol 62 (12) ◽

pp. 3594-3595 ◽

Cited By ~ 1

Author(s):

James R. Conner ◽

Robert H. Riddell

Keyword(s):

Colorectal Cancer ◽

High Grade Dysplasia ◽

Low Grade ◽

High Grade

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Stem-Like Signature Predicting Disease Progression in Early Stage Bladder Cancer. The Role of E2F3 and SOX4

Biomedicines ◽

10.3390/biomedicines6030085 ◽

2018 ◽

Vol 6 (3) ◽

pp. 85 ◽

Cited By ~ 4

Author(s):

Joaquim Bellmunt

Keyword(s):

Stem Cells ◽

Bladder Cancer ◽

Cancer Stem Cells ◽

Tumor Progression ◽

Cell Types ◽

Invasive Bladder Cancer ◽

Low Grade ◽

High Grade ◽

Muscle Invasive Bladder Cancer ◽

Muscle Invasive

The rapid development of the cancer stem cells (CSC) field, together with powerful genome-wide screening techniques, have provided the basis for the development of future alternative and reliable therapies aimed at targeting tumor-initiating cell populations. Urothelial bladder cancer stem cells (BCSCs) that were identified for the first time in 2009 are heterogenous and originate from multiple cell types; including urothelial stem cells and differentiated cell types—basal, intermediate stratum and umbrella cells Some studies hypothesize that BCSCs do not necessarily arise from normal stem cells but might derive from differentiated progenies following mutational insults and acquisition of tumorigenic properties. Conversely, there is data that normal bladder tissues can generate CSCs through mutations. Prognostic risk stratification by identification of predictive markers is of major importance in the management of urothelial cell carcinoma (UCC) patients. Several stem cell markers have been linked to recurrence or progression. The CD44v8-10 to standard CD44-ratio (total ratio of all CD44 alternative splicing isoforms) in urothelial cancer has been shown to be closely associated with tumor progression and aggressiveness. ALDH1, has also been reported to be associated with BCSCs and a worse prognosis in a large number of studies. UCC include low-grade and high-grade non-muscle invasive bladder cancer (NMIBC) and high-grade muscle invasive bladder cancer (MIBC). Important genetic defects characterize the distinct pathways in each one of the stages and probably grades. As an example, amplification of chromosome 6p22 is one of the most frequent changes seen in MIBC and might act as an early event in tumor progression. Interestingly, among NMIBC there is a much higher rate of amplification in high-grade NMIBC compared to low grade NMIBC. CDKAL1, E2F3 and SOX4 are highly expressed in patients with the chromosomal 6p22 amplification aside from other six well known genes (ID4, MBOAT1, LINC00340, PRL, and HDGFL1). Based on that, SOX4, E2F3 or 6q22.3 amplifications might represent potential targets in this tumor type. Focusing more in SOX4, it seems to exert its critical regulatory functions upstream of the Snail, Zeb, and Twist family of transcriptional inducers of EMT (epithelial–mesenchymal transition), but without directly affecting their expression as seen in several cell lines of the Cancer Cell Line Encyclopedia (CCLE) project. SOX4 gene expression correlates with advanced cancer stages and poor survival rate in bladder cancer, supporting a potential role as a regulator of the bladder CSC properties. SOX4 might serve as a biomarker of the aggressive phenotype, also underlying progression from NMIBC to MIBC. The amplicon in chromosome 6 contains SOX4 and E2F3 and is frequently found amplified in bladder cancer. These genes/amplicons might be a potential target for therapy. As an existing hypothesis is that chromatin deregulation through enhancers or super-enhancers might be the underlying mechanism responsible of this deregulation, a potential way to target these transcription factors could be through epigenetic modifiers.

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A phase II trial of maintenance ADAPT therapy targeting colon cancer stem cells in patients with metastatic colorectal cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.15_suppl.tps3650 ◽

2014 ◽

Vol 32 (15_suppl) ◽

pp. TPS3650-TPS3650 ◽

Cited By ~ 2

Author(s):

Edward H. Lin ◽

Shilpen A. Patel ◽

Jeffrey Chou ◽

Edward Y. Kim ◽

Veena Shankaran ◽

...

Keyword(s):

Colorectal Cancer ◽

Stem Cells ◽

Colon Cancer ◽

Cancer Stem Cells ◽

Metastatic Colorectal Cancer ◽

Phase Ii ◽

Phase Ii Trial ◽

Colon Cancer Stem Cells

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Human Colon Cancer Stem Cells: A New Paradigm in Gastrointestinal Oncology

Journal of Clinical Oncology ◽

10.1200/jco.2008.17.6941 ◽

2008 ◽

Vol 26 (17) ◽

pp. 2828-2838 ◽

Cited By ~ 121

Author(s):

Bruce M. Boman ◽

Emina Huang

Keyword(s):

Colorectal Cancer ◽

Stem Cells ◽

Colon Cancer ◽

Cancer Stem Cells ◽

Tumor Growth ◽

Human Colon ◽

Tissue Level ◽

New Paradigm ◽

Human Colon Cancer ◽

Recent Developments

For the past half century, oncologists have had systemic drugs available, agents that are able to induce tumor responses in patients with colorectal cancer. However, in cases of advanced colorectal cancer, these regimens are almost never curative. The recently introduced concept that cancer stem cells (SCs) drive tumor growth suggests a reason for these therapeutic failures— current chemotherapeutics target rapidly dividing cells but cancer SCs divide only slowly, and, they are relatively resistant to cytotoxic systemic therapies. It also suggests a solution—development of therapeutics that target cancer SCs. However, there is a paucity of information about the mechanisms by which SC populations are maintained and about the mechanisms by which tumor SCs are involved in colon cancer development. In this article, we discuss these mechanisms and recent developments in the identification and isolation of colon cancer SCs using new SC markers. We then discuss the role of SCs in homeostasis of normal colonic epithelium, and mechanisms by which dysregulation of crypt mechanisms can lead to initiation and progression of colon cancer. Our hypothesis, which has received recent experimental support, is that the mechanism that links abnormalities at the gene level (eg, APC mutations) and abnormalities at the tissue level (eg, proliferative shift, dysplasia, carcinoma) from cancer initiation to metastasis is SC overpopulation. Finally, we discuss the concept that symmetric cancer SC division is an essential mechanism that drives tumor growth, and that development of a new generation of therapeutics that target colon cancer SCs by inhibiting symmetric SC division holds promise for truly curative approaches for patients with advanced colorectal cancers.

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