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

Abstract

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

Cancer Stem-like Properties in Colorectal Cancer Cells with Low Proteasome Activity

“…Several CSC markers have been reported in colorectal cancer (7)(8)(9), and the population of CSCs in colorectal cancer has been reported to be 1.6% to 8.1% (8,36). On the other hand, in the field of hematologic malignancies, in which CSCs have been well studied, the population of CSCs has been reported to be <1% (35,37).…”

“…We used FACSAria II (BD Biosciences) for cell sorting and FACSDiva software (BD Biosciences) for analysis. CD24, CD26, CD44, CD133, and CD166 were previously reported to be typical markers of CSCs in colorectal cancer (7)(8)(9). CD24 (PE/Cy7 channel; Biolegend), CD26 (PE channel; BD Biosciences), CD44 (Alexa Fluor 700 channel; BD Biosciences), CD133 (APC channel; Miltenyi Biotec), and CD166 (PE channel; BD Biosciences) were used for the analysis of CSC markers according to the manufacturers' instructions.…”

“…5). Using similar methodology, several markers have been reported for the identification of CSCs in solid cancer, depending on the type of cancer: CD44 þ /CD24 À/low in breast cancer (6); CD24, CD26, CD44, CD133, and CD166 in colorectal cancer (7)(8)(9); CD24, CD44, and ESA in pancreatic cancer (10); and CD44, CD133, CD90, and ESA/EpCAM in hepatocellular carcinoma (11).…”

Targeted Confocal Laser Endomicroscopy (CLE) for the Assessment of Putative Cancer Stem Cell Markers in Colorectal cancer - A Pilot Study

“…Two other CSCs biomarkers (CD133 and CD166) were tested for evaluating their CLE expression but without relevant results. Improvements might be required regarding the CLE immunostaining technique, taking into consideration one of our previous histology study centered on CD133 and CD166 putative stem cells antibodies which registered consistent outcomes (52).…”

Revolutionizing the landscape of colorectal cancer treatment: The potential role of immune checkpoint inhibitors

“…76 Retrospective data indicated that CD166/CD133 colocalization was correlated to higher grade dysplasia in colon adenocarcinoma. 77 Moreover, the positive expression of CD166/CD44 in association with KRAS mutation is considered a poor prognostic feature linked to lymph node involvement as well as distant metastasis in patients with CRC. 78 CX-2009 is a novel CD166 probody conjugated with the tubulinbinding agent DM4.…”