Expression of the Stem Cell Markers Nestin and CD133 on Circulating Melanoma Cells

Abstract

Different molecular markers have been identified for melanoma-initiating cells including CD133 and nestin. Assuming that metastasis requires a dissemination of tumor-initiating cells, presence of circulating tumor-initiating cells should be associated with worse patient outcome. In this study, 20 ml blood was collected from 32 consecutive patients affected by metastatic melanoma and blood was enriched for circulating melanoma cells (CMCs) by CD45 depletion of the non-melanoma cell fraction. Multiparameter cytometry was carried out to co-stain with combinations of CD133 and nestin (NES). Six tissue samples from metastatic lesions of six different patients were stained with the same antibodies by immunohistochemistry. Percentage of NES-positive CMCs correlated with tumor burden and number of metastatic sites. Cox regression analysis revealed levels of lactate dehydrogenase (LDH; hazard ratio: 12.8 (1.35-121.5); P=0.02), number of metastatic sites (hazard ratio 3.87 (1.66-9.03); P=0.02), tumor burden (hazard ratio 5.72 (1.57-20.9); P=0.01), and percentage of NES-expressing CMCs ≥ 35% (hazard ratio 5.73 (1.66-19.7); P=0.006) to be factors related to shorter overall survival. CD133- and NES-expression profiles on CMCs were similar to matched metastatic tissue. These findings show that CMCs expressed stem cell-associated markers NES and CD133. Higher expression of NES on CMCs might represent an index of poor prognosis.

Co-Expression of Putative Cancer Stem Cell Markers, CD133 and Nestin, in Skin Tumors

“…The expression of Nestin by melanoma tumor cells was also reported to be associated with melanoma invasiveness (Tanabe et al, 2010). Recent findings suggest that CD133 and Nestin can be used as putative cancer stem cell markers in malignant melanoma (Klein et al, 2006;Monzani et al, 2007;Fusi et al, 2011;Thill et al, 2011).…”

“…In another study, Circulating Melanoma Cells (CMCs) were evaluated for the expression of CD133 and Nestin suggesting that CMCs expressed both markers, while higher expression of Nestin on CMCs have been found to be an index of poor prognosis (Fusi et al, 2011).…”

“…The expression of Nestin in melanoma tumor cells was also correlated with melanoma invasiveness (Tanabe et al, 2010). Moreover, Nestin was detected in the blood of the patients with melanoma (Fusi et al, 2011), and its detection was associated with the presence of circulating melanoma tumor cells indicating the metastatic tendency of melanoma. Therefore, Nestin might be an early biomarker for melanoma recurrence (Fusi et al, 2011).…”

“…CSCs have been identified in a number of cancers including acute myeloid leukaemia (AML) (Bonnet and Dick, 1997;Passegue et al, 2003), glioblastoma (Singh et al, 2003), breast (Al-Hajj et al, 2003;Ponti et al, 2005), lung (Kim et al, 2005), prostate (Collins et al, 2005), ovarian (Bapat et al, 2005), gastric (Houghton et al, 2004), esophagous ( Li et al, 2013 ), Head and Neck SCC (Satpute et al, 2013) and skin cancers (Frank et al, 2005;Monzani et al, 2007). Different markers have been identified to be expressed on melanoma stem cells (Quintana et al, 2010;Shakhova and Sommer, 2013) comprising CD20 (Fang et al, 2005) and ABC transporter family members such as MDR1, ABCG2 and ABCB5 (Frank et al, 2003;Frank et al, 2005;Monzani et al, 2007;Keshet et al, 2008;Schatton et al, 2008), CD271 (Boiko et al, 2010;Civenni et al, 2011), CD44 (Fernandez-Figueras et al, 1996, CD133 (Klein et al, 2006) and Nestin (Piras et al, 2010;Fusi et al, 2011). Among these markers, CD133 and Nestin have been described as markers of melanocytic stem cells and two of the most important surface markers with increased expression in the cancer stem cell fraction in different human malignancies, including melanoma (Klein et al, 2006;Monzani et al, 2007;Rappa et al, 2008;Al Dhaybi et al, 2010;Shakhova and Sommer, 2013).…”

Mixed versus pure variants of desmoplastic melanoma: a genetic and immunohistochemical appraisal

“…KIT was investigated on the basis of the rationale that melanomas lacking BRAF mutations may have mutations or copy number alterations in c-KIT. 12 Immunohistochemical markers included the following: nestin, an intermediate filament protein, a melanocyte stem cell marker and a marker for melanoma-initiating cells, associated with poor prognosis in conventional melanoma; 13 clusterin, an 80-kDa glycoprotein of undefined biological significance shown to be significantly increased in desmoplastic melanoma; 11 SOX10, a neural crest stem cell marker, expressed in melanocytes, specifically, strongly expressed by desmoplastic melanomas; 14 CD117, a stem cell growth factor receptor encoded by the KIT gene, a receptor tyrosine kinase involved in cell survival and proliferation of melanocytes; CD271 (p75NTR), a neural crest stem cell factor, which has been shown to correlate with higher metastatic potential and worse clinical outcomes in melanoma; 15 and Ki-67 protein, a marker of cellular proliferation, which is expressed during of all phases of cell cycling (G1, S, G2 and mitosis), but absent during the resting phase. 16 …”