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Effect of MUC1/ β -catenin interaction on the tumorigenic capacity of pancreatic CD133 + cells

View Article: PubMed Central - PubMed

ABSTRACT

Despite the fact that the biological function of cluster of differentiation (CD)133 remains unclear, this glycoprotein is currently used in the identification and isolation of tumor-initiating cells from certain malignant tumors, including pancreatic cancer. In the present study, the involvement of mucin 1 (MUC1) in the signaling pathways of a highly tumorigenic CD133+ cellular subpopulation sorted from the pancreatic cancer cell line HPAF-II was evaluated. The expression of MUC1-cytoplasmic domain (MUC1-CD) and oncogenic signaling transducers (epidermal growth factor receptor, protein kinase C delta, glycogen synthase kinase 3 beta and growth factor receptor-bound protein 2), as well as the association between MUC1 and β-catenin, were characterized in HPAF-II CD133+ and CD133low cell subpopulations and in tumor xenografts generated from these cells. Compared with HPAF CD133low cells, HPAF-II CD133+ cancer cells exhibited increased tumorigenic potential in immunocompromised mice, which was associated with overexpression of MUC1 and with the accordingly altered expression profile of MUC1-associated signaling partners. Additionally, MUC1-CD/β-catenin interactions were increased both in the HPAF-II CD133+ cell subpopulation and derived tumor xenografts compared with HPAF CD133low cells. These results suggest that, in comparison with HPAF CD133low cells, CD133+ cells exhibit higher expression of MUC1, which contributes to their tumorigenic phenotype through increased interaction between MUC1-CD and β-catenin, which in turn modulates oncogenic signaling cascades.

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Validation of the CSC model. (A) Identification of a CSC subpopulation (CD133+ cells) in the HPAF-II pancreatic cancer cell line and evaluation of CD133 expression in cell subpopulations sorted by flow cytometry. (a) Isotype stained cells were used as controls. (b) HPAF-II cells stained with CD133/2-phycoerythrin monoclonal antibody. (B) Enrichment of HPAF-II CD133+ subpopulation isolated by magnetic-activated cell sorting represented on a frequency distribution histogram. The HPAF-II CD133+ subpopulation exhibits 8.89% of CD133+ cells, while the HPAF-II CD133low subpopulation exhibited 3.07% of CD133+ cells, representing an enriched and a depleted population, respectively. (C) CD133 expression in HPAF-II tumor xenografts determined by immunohistochemistry (magnification, ×400). Hemotoxylin and eosin staining was used to reveal the morphology of the tumors (magnification, ×100). CSC, cancer stem cell; CD, cluster of differentiation; H&E, hematoxylin and eosin; SSC-H, measures cell granularity or internal complexity; FH2, phycoerythrin detection; % of Max, % of maximum (normalization method).
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f1-ol-0-0-4888: Validation of the CSC model. (A) Identification of a CSC subpopulation (CD133+ cells) in the HPAF-II pancreatic cancer cell line and evaluation of CD133 expression in cell subpopulations sorted by flow cytometry. (a) Isotype stained cells were used as controls. (b) HPAF-II cells stained with CD133/2-phycoerythrin monoclonal antibody. (B) Enrichment of HPAF-II CD133+ subpopulation isolated by magnetic-activated cell sorting represented on a frequency distribution histogram. The HPAF-II CD133+ subpopulation exhibits 8.89% of CD133+ cells, while the HPAF-II CD133low subpopulation exhibited 3.07% of CD133+ cells, representing an enriched and a depleted population, respectively. (C) CD133 expression in HPAF-II tumor xenografts determined by immunohistochemistry (magnification, ×400). Hemotoxylin and eosin staining was used to reveal the morphology of the tumors (magnification, ×100). CSC, cancer stem cell; CD, cluster of differentiation; H&E, hematoxylin and eosin; SSC-H, measures cell granularity or internal complexity; FH2, phycoerythrin detection; % of Max, % of maximum (normalization method).

Mentions: Low-passage/highly tumorigenic samples of the HPAF-II cell line (104 cells produced tumors in 100% of animals; data not shown) were evaluated for CD133 expression levels by flow cytometry. The results indicated that low-passage HPAF-II cells contained ~4% CD133+ cells (Fig. 1A). These cells were isolated using MACS, and both CD133+ and CD133− subpopulations were cultured. To evaluate the enrichment obtained with the sorting methodology used, CD133 was again measured by flow cytometry in the above two cell subpopulations prior to injection into immunodeficient mice (Fig. 1B). The results revealed that the CD133+ subpopulation was highly enriched in CD133+ cells. However, the putative CD133− subpopulation retained a very low percentage of cells expressing CD133, and was therefore called CD133low. Repeated selection did not improve the performance of this procedure (data not shown).


Effect of MUC1/ β -catenin interaction on the tumorigenic capacity of pancreatic CD133 + cells
Validation of the CSC model. (A) Identification of a CSC subpopulation (CD133+ cells) in the HPAF-II pancreatic cancer cell line and evaluation of CD133 expression in cell subpopulations sorted by flow cytometry. (a) Isotype stained cells were used as controls. (b) HPAF-II cells stained with CD133/2-phycoerythrin monoclonal antibody. (B) Enrichment of HPAF-II CD133+ subpopulation isolated by magnetic-activated cell sorting represented on a frequency distribution histogram. The HPAF-II CD133+ subpopulation exhibits 8.89% of CD133+ cells, while the HPAF-II CD133low subpopulation exhibited 3.07% of CD133+ cells, representing an enriched and a depleted population, respectively. (C) CD133 expression in HPAF-II tumor xenografts determined by immunohistochemistry (magnification, ×400). Hemotoxylin and eosin staining was used to reveal the morphology of the tumors (magnification, ×100). CSC, cancer stem cell; CD, cluster of differentiation; H&E, hematoxylin and eosin; SSC-H, measures cell granularity or internal complexity; FH2, phycoerythrin detection; % of Max, % of maximum (normalization method).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4998183&req=5

f1-ol-0-0-4888: Validation of the CSC model. (A) Identification of a CSC subpopulation (CD133+ cells) in the HPAF-II pancreatic cancer cell line and evaluation of CD133 expression in cell subpopulations sorted by flow cytometry. (a) Isotype stained cells were used as controls. (b) HPAF-II cells stained with CD133/2-phycoerythrin monoclonal antibody. (B) Enrichment of HPAF-II CD133+ subpopulation isolated by magnetic-activated cell sorting represented on a frequency distribution histogram. The HPAF-II CD133+ subpopulation exhibits 8.89% of CD133+ cells, while the HPAF-II CD133low subpopulation exhibited 3.07% of CD133+ cells, representing an enriched and a depleted population, respectively. (C) CD133 expression in HPAF-II tumor xenografts determined by immunohistochemistry (magnification, ×400). Hemotoxylin and eosin staining was used to reveal the morphology of the tumors (magnification, ×100). CSC, cancer stem cell; CD, cluster of differentiation; H&E, hematoxylin and eosin; SSC-H, measures cell granularity or internal complexity; FH2, phycoerythrin detection; % of Max, % of maximum (normalization method).
Mentions: Low-passage/highly tumorigenic samples of the HPAF-II cell line (104 cells produced tumors in 100% of animals; data not shown) were evaluated for CD133 expression levels by flow cytometry. The results indicated that low-passage HPAF-II cells contained ~4% CD133+ cells (Fig. 1A). These cells were isolated using MACS, and both CD133+ and CD133− subpopulations were cultured. To evaluate the enrichment obtained with the sorting methodology used, CD133 was again measured by flow cytometry in the above two cell subpopulations prior to injection into immunodeficient mice (Fig. 1B). The results revealed that the CD133+ subpopulation was highly enriched in CD133+ cells. However, the putative CD133− subpopulation retained a very low percentage of cells expressing CD133, and was therefore called CD133low. Repeated selection did not improve the performance of this procedure (data not shown).

View Article: PubMed Central - PubMed

ABSTRACT

Despite the fact that the biological function of cluster of differentiation (CD)133 remains unclear, this glycoprotein is currently used in the identification and isolation of tumor-initiating cells from certain malignant tumors, including pancreatic cancer. In the present study, the involvement of mucin 1 (MUC1) in the signaling pathways of a highly tumorigenic CD133+ cellular subpopulation sorted from the pancreatic cancer cell line HPAF-II was evaluated. The expression of MUC1-cytoplasmic domain (MUC1-CD) and oncogenic signaling transducers (epidermal growth factor receptor, protein kinase C delta, glycogen synthase kinase 3 beta and growth factor receptor-bound protein 2), as well as the association between MUC1 and β-catenin, were characterized in HPAF-II CD133+ and CD133low cell subpopulations and in tumor xenografts generated from these cells. Compared with HPAF CD133low cells, HPAF-II CD133+ cancer cells exhibited increased tumorigenic potential in immunocompromised mice, which was associated with overexpression of MUC1 and with the accordingly altered expression profile of MUC1-associated signaling partners. Additionally, MUC1-CD/β-catenin interactions were increased both in the HPAF-II CD133+ cell subpopulation and derived tumor xenografts compared with HPAF CD133low cells. These results suggest that, in comparison with HPAF CD133low cells, CD133+ cells exhibit higher expression of MUC1, which contributes to their tumorigenic phenotype through increased interaction between MUC1-CD and β-catenin, which in turn modulates oncogenic signaling cascades.

No MeSH data available.


Related in: MedlinePlus