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Characterization of colon cancer cells: a functional approach characterizing CD133 as a potential stem cell marker.

Schneider M, Huber J, Hadaschik B, Siegers GM, Fiebig HH, Schüler J - BMC Cancer (2012)

Bottom Line: In 8/14 cell lines, more than 80% of the cells were positive for CD24 and 11/14 were over 70% positive for CD44. 10/14 cell lines expressed CDCP1 on ≥ 83% of cells.Our data revealed correlations in the expression of surface markers CD44 and CD24 as well as CD44 and CDCP1 and strongly suggest that CD133 is a stem cell marker within our colon carcinoma panel.Further studies will elucidate its role as a potential therapeutic target.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Urology, University of Heidelberg, Im Neuenheimer Feld 110, D-69120 Heidelberg, Germany. meike.schneider@med.uni-heidelberg.de

ABSTRACT

Background: Isolation and characterization of tumourigenic colon cancer initiating cells may help to develop novel diagnostic and therapeutic procedures.

Methods: We characterized a panel of fourteen human colon carcinoma cell lines and their corresponding xenografts for the surface expression of potential stem cell markers CD133, CD24, CD44, CDCP1 and CXCR4. In five cell lines and nine xenografts, mRNA expression of these markers was determined. Tumour growth behaviour of CD133+, CD133- and unsorted SW620 cells was evaluated in vivo.

Results: All five putative stem cell markers showed distinct expression patterns in the tumours examined. Two patient-derived cell lines highly expressed CD133 (> 85% of positive cells) and three other cell lines had an expression level of about 50% whereas in long-term culture based models CD133 expression ranged only from 0 to 20%. In 8/14 cell lines, more than 80% of the cells were positive for CD24 and 11/14 were over 70% positive for CD44. 10/14 cell lines expressed CDCP1 on ≥ 83% of cells. CXCR4 expression was determined solely on 94 L and SW480.Analyses of the corresponding xenografts revealed a significant reduction of cell numbers expressing the investigated surface markers and showed single cell fractions expressing up to three markers simultaneously.Statistical analysis revealed that the CXCR4 mRNA level correlates negatively with the protein expression of CD133, CD44, CD24 and CDCP1 in cell lines and xenografts.A lower differentiation grade of donor material correlated with a higher CDCP1 mRNA expression level in the respective tumour model.In vivo growth behaviour studies of SW620 revealed significantly higher take rates and shorter doubling times in the tumour growth of CD133 positive subclones in comparison to the unsorted cell line or CD133 negative subclones.

Conclusions: Our data revealed correlations in the expression of surface markers CD44 and CD24 as well as CD44 and CDCP1 and strongly suggest that CD133 is a stem cell marker within our colon carcinoma panel. Further studies will elucidate its role as a potential therapeutic target.

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Expression of 5 different surface markers on a panel of human colon carcinoma xenografts.
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Figure 3: Expression of 5 different surface markers on a panel of human colon carcinoma xenografts.

Mentions: Mean expression of CD133 on cell lines was 23.5 ± 15.2% and 14.5 ± 7.2% on xenografts (Figure 1). With respect to CD133 expression, the cell lines could be subdivided into four groups: five cell lines had a very small subpopulation of CD133 expressing cells (≤ 0.02%); four cell lines expressed 8-15% CD133; in CCL HT29, LOVO and SW620, 50% of the cells expressed CD133; the highest CD133 expression levels were found on Oncotest proprietary cell lines 269 L and 94 L (85.5 and 85.8% respectively) (Figure 2). Examining the colon xenografts, four models revealed very weak CD133 expression ranging between 0 and 0.2%. A group of six tumours expressed CD133 ranging from 4.0 to 14.5%. Another five had expression levels of 20.7 to 36.6% (Figure 3).


Characterization of colon cancer cells: a functional approach characterizing CD133 as a potential stem cell marker.

Schneider M, Huber J, Hadaschik B, Siegers GM, Fiebig HH, Schüler J - BMC Cancer (2012)

Expression of 5 different surface markers on a panel of human colon carcinoma xenografts.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3368744&req=5

Figure 3: Expression of 5 different surface markers on a panel of human colon carcinoma xenografts.
Mentions: Mean expression of CD133 on cell lines was 23.5 ± 15.2% and 14.5 ± 7.2% on xenografts (Figure 1). With respect to CD133 expression, the cell lines could be subdivided into four groups: five cell lines had a very small subpopulation of CD133 expressing cells (≤ 0.02%); four cell lines expressed 8-15% CD133; in CCL HT29, LOVO and SW620, 50% of the cells expressed CD133; the highest CD133 expression levels were found on Oncotest proprietary cell lines 269 L and 94 L (85.5 and 85.8% respectively) (Figure 2). Examining the colon xenografts, four models revealed very weak CD133 expression ranging between 0 and 0.2%. A group of six tumours expressed CD133 ranging from 4.0 to 14.5%. Another five had expression levels of 20.7 to 36.6% (Figure 3).

Bottom Line: In 8/14 cell lines, more than 80% of the cells were positive for CD24 and 11/14 were over 70% positive for CD44. 10/14 cell lines expressed CDCP1 on ≥ 83% of cells.Our data revealed correlations in the expression of surface markers CD44 and CD24 as well as CD44 and CDCP1 and strongly suggest that CD133 is a stem cell marker within our colon carcinoma panel.Further studies will elucidate its role as a potential therapeutic target.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Urology, University of Heidelberg, Im Neuenheimer Feld 110, D-69120 Heidelberg, Germany. meike.schneider@med.uni-heidelberg.de

ABSTRACT

Background: Isolation and characterization of tumourigenic colon cancer initiating cells may help to develop novel diagnostic and therapeutic procedures.

Methods: We characterized a panel of fourteen human colon carcinoma cell lines and their corresponding xenografts for the surface expression of potential stem cell markers CD133, CD24, CD44, CDCP1 and CXCR4. In five cell lines and nine xenografts, mRNA expression of these markers was determined. Tumour growth behaviour of CD133+, CD133- and unsorted SW620 cells was evaluated in vivo.

Results: All five putative stem cell markers showed distinct expression patterns in the tumours examined. Two patient-derived cell lines highly expressed CD133 (> 85% of positive cells) and three other cell lines had an expression level of about 50% whereas in long-term culture based models CD133 expression ranged only from 0 to 20%. In 8/14 cell lines, more than 80% of the cells were positive for CD24 and 11/14 were over 70% positive for CD44. 10/14 cell lines expressed CDCP1 on ≥ 83% of cells. CXCR4 expression was determined solely on 94 L and SW480.Analyses of the corresponding xenografts revealed a significant reduction of cell numbers expressing the investigated surface markers and showed single cell fractions expressing up to three markers simultaneously.Statistical analysis revealed that the CXCR4 mRNA level correlates negatively with the protein expression of CD133, CD44, CD24 and CDCP1 in cell lines and xenografts.A lower differentiation grade of donor material correlated with a higher CDCP1 mRNA expression level in the respective tumour model.In vivo growth behaviour studies of SW620 revealed significantly higher take rates and shorter doubling times in the tumour growth of CD133 positive subclones in comparison to the unsorted cell line or CD133 negative subclones.

Conclusions: Our data revealed correlations in the expression of surface markers CD44 and CD24 as well as CD44 and CDCP1 and strongly suggest that CD133 is a stem cell marker within our colon carcinoma panel. Further studies will elucidate its role as a potential therapeutic target.

Show MeSH
Related in: MedlinePlus