Limits...
Atypical nuclear localization of CD133 plasma membrane glycoprotein in rhabdomyosarcoma cell lines.

Nunukova A, Neradil J, Skoda J, Jaros J, Hampl A, Sterba J, Veselska R - Int. J. Mol. Med. (2015)

Bottom Line: In the present study, we verified the nuclear localization of CD133 in RMS cells using three independent anti-CD133 antibodies, including both rabbit polyclonal and mouse monoclonal antibodies.Indirect immunofluorescence and confocal microscopy followed by software cross-section analysis, transmission electron microscopy and cell fractionation with immunoblotting were also employed, and all the results undeniably confirmed the presence of CD133 in the nuclei of stable minor subpopulations of all five RMS cell lines.The proportion of cells showing an exclusive nuclear localization of CD133 ranged from 3.4 to 7.5%, with only minor differences observed among the individual anti-CD133 antibodies.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Biology, Faculty of Science, Masaryk University, 61137 Brno, Czech Republic.

ABSTRACT
CD133 (also known as prominin-1) is a cell surface glycoprotein that is widely used for the identification of stem cells. Furthermore, its glycosylated epitope, AC133, has recently been discussed as a marker of cancer stem cells in various human malignancies. During our recent experiments on rhabdomyosarcomas (RMS), we unexpectedly identified an atypical nuclear localization of CD133 in a relatively stable subset of cells in five RMS cell lines established in our laboratory. To the best of our knowledge, this atypical localization of CD133 has not yet been proven or analyzed in detail in cancer cells. In the present study, we verified the nuclear localization of CD133 in RMS cells using three independent anti-CD133 antibodies, including both rabbit polyclonal and mouse monoclonal antibodies. Indirect immunofluorescence and confocal microscopy followed by software cross-section analysis, transmission electron microscopy and cell fractionation with immunoblotting were also employed, and all the results undeniably confirmed the presence of CD133 in the nuclei of stable minor subpopulations of all five RMS cell lines. The proportion of cells showing an exclusive nuclear localization of CD133 ranged from 3.4 to 7.5%, with only minor differences observed among the individual anti-CD133 antibodies. Although the role of CD133 in the cell nucleus remains unclear, these results clearly indicate that this atypical nuclear localization of CD133 in a minor subpopulation of cancer cells is a common phenomenon in RMS cell lines.

No MeSH data available.


Related in: MedlinePlus

Overview of epitopes of the anti-CD133 and anti-AC133 antibodies used in this study. For each antibody, the catalogue number and manufacturer are indicated. Potential glycosylation sites, as well as length of the N-terminal region, the intracellular and extracellular loops and the C-terminal region of CD133 are depicted.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4494592&req=5

f1-ijmm-36-01-0065: Overview of epitopes of the anti-CD133 and anti-AC133 antibodies used in this study. For each antibody, the catalogue number and manufacturer are indicated. Potential glycosylation sites, as well as length of the N-terminal region, the intracellular and extracellular loops and the C-terminal region of CD133 are depicted.

Mentions: During our recent study aimed at the analysis of CSC markers in pediatric sarcomas, we noted a surprising result: a stable subset of cells in each of five RMS cell lines examined exhibited an exclusive nuclear localization of CD133 (these data are published in this article). To date, a similar localization of this antigen has been described only in one case report of breast cancer (14) and in a large study on lung cancer (15) using immunohistochemical methods, nevertheless, without any verification or systematic description. For this reason, in this study, we sought to analyze this interesting phenomenon in detail using three independent anti-CD133 commercial antibodies (Fig. 1).


Atypical nuclear localization of CD133 plasma membrane glycoprotein in rhabdomyosarcoma cell lines.

Nunukova A, Neradil J, Skoda J, Jaros J, Hampl A, Sterba J, Veselska R - Int. J. Mol. Med. (2015)

Overview of epitopes of the anti-CD133 and anti-AC133 antibodies used in this study. For each antibody, the catalogue number and manufacturer are indicated. Potential glycosylation sites, as well as length of the N-terminal region, the intracellular and extracellular loops and the C-terminal region of CD133 are depicted.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494592&req=5

f1-ijmm-36-01-0065: Overview of epitopes of the anti-CD133 and anti-AC133 antibodies used in this study. For each antibody, the catalogue number and manufacturer are indicated. Potential glycosylation sites, as well as length of the N-terminal region, the intracellular and extracellular loops and the C-terminal region of CD133 are depicted.
Mentions: During our recent study aimed at the analysis of CSC markers in pediatric sarcomas, we noted a surprising result: a stable subset of cells in each of five RMS cell lines examined exhibited an exclusive nuclear localization of CD133 (these data are published in this article). To date, a similar localization of this antigen has been described only in one case report of breast cancer (14) and in a large study on lung cancer (15) using immunohistochemical methods, nevertheless, without any verification or systematic description. For this reason, in this study, we sought to analyze this interesting phenomenon in detail using three independent anti-CD133 commercial antibodies (Fig. 1).

Bottom Line: In the present study, we verified the nuclear localization of CD133 in RMS cells using three independent anti-CD133 antibodies, including both rabbit polyclonal and mouse monoclonal antibodies.Indirect immunofluorescence and confocal microscopy followed by software cross-section analysis, transmission electron microscopy and cell fractionation with immunoblotting were also employed, and all the results undeniably confirmed the presence of CD133 in the nuclei of stable minor subpopulations of all five RMS cell lines.The proportion of cells showing an exclusive nuclear localization of CD133 ranged from 3.4 to 7.5%, with only minor differences observed among the individual anti-CD133 antibodies.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Biology, Faculty of Science, Masaryk University, 61137 Brno, Czech Republic.

ABSTRACT
CD133 (also known as prominin-1) is a cell surface glycoprotein that is widely used for the identification of stem cells. Furthermore, its glycosylated epitope, AC133, has recently been discussed as a marker of cancer stem cells in various human malignancies. During our recent experiments on rhabdomyosarcomas (RMS), we unexpectedly identified an atypical nuclear localization of CD133 in a relatively stable subset of cells in five RMS cell lines established in our laboratory. To the best of our knowledge, this atypical localization of CD133 has not yet been proven or analyzed in detail in cancer cells. In the present study, we verified the nuclear localization of CD133 in RMS cells using three independent anti-CD133 antibodies, including both rabbit polyclonal and mouse monoclonal antibodies. Indirect immunofluorescence and confocal microscopy followed by software cross-section analysis, transmission electron microscopy and cell fractionation with immunoblotting were also employed, and all the results undeniably confirmed the presence of CD133 in the nuclei of stable minor subpopulations of all five RMS cell lines. The proportion of cells showing an exclusive nuclear localization of CD133 ranged from 3.4 to 7.5%, with only minor differences observed among the individual anti-CD133 antibodies. Although the role of CD133 in the cell nucleus remains unclear, these results clearly indicate that this atypical nuclear localization of CD133 in a minor subpopulation of cancer cells is a common phenomenon in RMS cell lines.

No MeSH data available.


Related in: MedlinePlus