Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2008 Oct 1;68(19):7882-6.
doi: 10.1158/0008-5472.CAN-08-0723.

Cell cycle-dependent variation of a CD133 epitope in human embryonic stem cell, colon cancer, and melanoma cell lines

Marie Jaksch  1 , Jorge Múnera, Ruchi Bajpai, Alexey Terskikh, Robert G Oshima
Affiliations

Cell cycle-dependent variation of a CD133 epitope in human embryonic stem cell, colon cancer, and melanoma cell lines

Marie Jaksch et al. Cancer Res. .

Abstract

CD133 (Prominin1) is a pentaspan transmembrane glycoprotein expressed in several stem cell populations and cancers. Reactivity with an antibody (AC133) to a glycoslyated form of CD133 has been widely used for the enrichment of cells with tumor-initiating activity in xenograph transplantation assays. We have found by fluorescence-activated cell sorting that increased AC133 reactivity in human embryonic stem cells, colon cancer, and melanoma cells is correlated with increased DNA content and, reciprocally, that the least reactive cells are in the G(1)-G(0) portion of the cell cycle. Continued cultivation of cells sorted on the basis of high and low AC133 reactivity results in a normalization of the cell reactivity profiles, indicating that cells with low AC133 reactivity can generate highly reactive cells as they resume proliferation. The association of AC133 with actively cycling cells may contribute to the basis for enrichment for tumor-initiating activity.

PubMed Disclaimer

Figures

Figure 1
Figure 1. AC133 expression in Caco2 and hES cells
A, Immunofluorescent analysis of AC133 staining in Caco2 colon cancer cells and cytospin preparation of H9 hES cells (40× magnification, scale bar 100μM). B, Flow cytometry analysis of AC133 staining on live cells (negative controls in grey). C. AC133 expression on confluent, sub-confluent and post-confluent Caco2 cells.
Figure 2
Figure 2. Correlation of AC133 reaction and DNA content
A, The AC133 reactive populations of hES cells were divided into five groups (1-5), each representing twenty percent of the total population. B, DNA contents for the individual fractions of AC133 reactive cells. Graph no. 6 shows the cell cycle profile for all AC133 positive cells. C, Graphs show the percentage of cells with 2N DNA content and cells with 4N DNA content for each subpopulation of cells with AC133 reactivity and cells with MELK-GFP expression. D, DNA content profiles for cells gated on the highest 10% (black line, no fill) and the lowest 10% (grey fill) of AC133 expression in three different cell lines, hES cells, melanoma cells WM115 and colon cancer Caco2 cells.
Figure 3
Figure 3. Gene expression in AC133 high vs. negative cells
A and B. Scatter plots of AC133 high vs. negative cells for Caco2 and WM115 cells, respectively. Fold change ≥ 2 are indicated above and below the lines parallel to the diagonal. Not all 2× changes are statistically significant. C. Cluster analysis of microarray data. Gene expression in AC133 high and negative sorted Caco2 (sample A-D) and WM115 (sample E-H). The Pearson correlation using a 1-r distance measure was applied.
Figure 4
Figure 4. Colony formation, proliferation and AC133 expression in AC133 high and negative sorted cells
A, Colony formation of AC133 high and negative sorted Caco2 cells, (colony forming units (cfu)/seeded cells). B, Proliferation rate in AC133 high and negative sorted Caco2 cells. C, show AC133 expression in sorted Caco2 cells before and after cultivation of negative and high sorted cells, respectively. The grey fill profiles show AC133 expression of the sorted cells and the black lines show AC133 expression after 3 passages of cultivation. D, Morphology of AC133 high and negative sorted Caco2 cells after 7 days of cultivation (20× magnification, scale bar 100μM).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Ailles LE, Weissman IL. Cancer stem cells in solid tumors. Curr Opin Biotechnol. 2007;18:460–6. - PubMed
    1. Beier D, Hau P, Proescholdt M, et al. CD133(+) and CD133(-) glioblastoma-derived cancer stem cells show differential growth characteristics and molecular profiles. Cancer Res. 2007;67:4010–5. - PubMed
    1. Shmelkov SV, Butler JM, Hooper AT, et al. CD133 expression is not restricted to stem cells, and both CD133+ and CD133- metastatic colon cancer cells initiate tumors. J Clin Invest. 2008;118:2111–20. - PMC - PubMed
    1. Zheng X, Shen G, Yang X, Liu W. Most C6 cells are cancer stem cells: evidence from clonal and population analyses. Cancer Res. 2007;67:3691–7. - PubMed
    1. Miraglia S, Godfrey W, Yin AH, et al. A novel five-transmembrane hematopoietic stem cell antigen: isolation, characterization, and molecular cloning. Blood. 1997;90:5013–21. - PubMed

Publication types

MeSH terms