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Heterogeneity of circulating epithelial tumour cells from individual patients with respect to expression profiles and clonal growth (sphere formation) in breast cancer.

Pizon M, Zimon D, Carl S, Pachmann U, Pachmann K, Camara O - Ecancermedicalscience (2013)

Bottom Line: Here we present data that a variable fraction among the circulating tumour cells detected by the Maintrac(®) approach expresses mRNA of the stem cell gene NANOG and of the adhesion molecule vimentin and is capable of forming tumour spheres, a property ascribed to tumour-initiating cells (TICs).Although no epithelial cell adhesion molecule (EpCAM)-positive cells expressing stem cell genes or the adhesion molecule vimentin was detected before surgery, 10%-20% of the cells were found to be positive for mRNA of these genes after surgery.Here we show that among the peripherally circulating tumour cells, a variable fraction is able to express stem cell and adhesion properties and can be grown into tumour spheres, a property ascribed to cells capable of initiating tumours and metastases.

View Article: PubMed Central - PubMed

Affiliation: Transfusion Center Bayreuth D-95448, Germany.

ABSTRACT

Background: The detection of tumour cells circulating in the peripheral blood of patients with breast cancer is a sign that cells have been able to leave the primary tumour and survive in the circulation. However, in order to form metastases, they require additional properties such as the ability to adhere, self-renew, and grow. Here we present data that a variable fraction among the circulating tumour cells detected by the Maintrac(®) approach expresses mRNA of the stem cell gene NANOG and of the adhesion molecule vimentin and is capable of forming tumour spheres, a property ascribed to tumour-initiating cells (TICs).

Patients and methods: Between ten and 50 circulating epithelial antigen-positive cells detected by the Maintrac approach were selected randomly from each of 20 patients with breast cancer before and after surgery and were isolated using automated capillary aspiration and deposited individually onto slides for expression profiling. In addition, the circulating tumour cells were cultured without isolation among the white blood cells from 39 patients with breast cancer in different stages of disease using culture methods favouring growth of epithelial cells.

Results: Although no epithelial cell adhesion molecule (EpCAM)-positive cells expressing stem cell genes or the adhesion molecule vimentin was detected before surgery, 10%-20% of the cells were found to be positive for mRNA of these genes after surgery. Tumour spheres from circulating cells of 39 patients with different stages of breast cancer were grown without previous isolation in a fraction increasing with the aggressivity of the tumour.

Summary: Here we show that among the peripherally circulating tumour cells, a variable fraction is able to express stem cell and adhesion properties and can be grown into tumour spheres, a property ascribed to cells capable of initiating tumours and metastases.

No MeSH data available.


Related in: MedlinePlus

(a) Four vital epithelial antigen-positive tumour cells (only green fluorescent, thick arrows) and three avital (green and red fluorescent, thin arrows) among the unstained live and red fluorescent nuclei of avital blood cells. (b) Typical cells used for individual selection in transmitted light and green fluorescence (small pictures) and the merged pictures (large pictures). (c) The semiautomated capillary device for aspiration of individual cells from cell suspensions (slides in the front) and deposition in indiviual wells (slide in the background).
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figure1: (a) Four vital epithelial antigen-positive tumour cells (only green fluorescent, thick arrows) and three avital (green and red fluorescent, thin arrows) among the unstained live and red fluorescent nuclei of avital blood cells. (b) Typical cells used for individual selection in transmitted light and green fluorescence (small pictures) and the merged pictures (large pictures). (c) The semiautomated capillary device for aspiration of individual cells from cell suspensions (slides in the front) and deposition in indiviual wells (slide in the background).

Mentions: For expression analysis, vital cells stained with membrane epithelial cell adhesion molecule (EpCAM) and excluding propidium iodide (PI) as an avitality marker among the white blood cells from 20 breast cancer patients before and after surgery, comprising the tumour suspect cells from 1 mL of blood, were selected individually using the CellEctor Plus (MMI, Zurich, Switzerland) and deposited one by one onto Advalytix AmpliGrid Microscope Slides (Hamilton) (Figures 1a–c).


Heterogeneity of circulating epithelial tumour cells from individual patients with respect to expression profiles and clonal growth (sphere formation) in breast cancer.

Pizon M, Zimon D, Carl S, Pachmann U, Pachmann K, Camara O - Ecancermedicalscience (2013)

(a) Four vital epithelial antigen-positive tumour cells (only green fluorescent, thick arrows) and three avital (green and red fluorescent, thin arrows) among the unstained live and red fluorescent nuclei of avital blood cells. (b) Typical cells used for individual selection in transmitted light and green fluorescence (small pictures) and the merged pictures (large pictures). (c) The semiautomated capillary device for aspiration of individual cells from cell suspensions (slides in the front) and deposition in indiviual wells (slide in the background).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750196&req=5

figure1: (a) Four vital epithelial antigen-positive tumour cells (only green fluorescent, thick arrows) and three avital (green and red fluorescent, thin arrows) among the unstained live and red fluorescent nuclei of avital blood cells. (b) Typical cells used for individual selection in transmitted light and green fluorescence (small pictures) and the merged pictures (large pictures). (c) The semiautomated capillary device for aspiration of individual cells from cell suspensions (slides in the front) and deposition in indiviual wells (slide in the background).
Mentions: For expression analysis, vital cells stained with membrane epithelial cell adhesion molecule (EpCAM) and excluding propidium iodide (PI) as an avitality marker among the white blood cells from 20 breast cancer patients before and after surgery, comprising the tumour suspect cells from 1 mL of blood, were selected individually using the CellEctor Plus (MMI, Zurich, Switzerland) and deposited one by one onto Advalytix AmpliGrid Microscope Slides (Hamilton) (Figures 1a–c).

Bottom Line: Here we present data that a variable fraction among the circulating tumour cells detected by the Maintrac(®) approach expresses mRNA of the stem cell gene NANOG and of the adhesion molecule vimentin and is capable of forming tumour spheres, a property ascribed to tumour-initiating cells (TICs).Although no epithelial cell adhesion molecule (EpCAM)-positive cells expressing stem cell genes or the adhesion molecule vimentin was detected before surgery, 10%-20% of the cells were found to be positive for mRNA of these genes after surgery.Here we show that among the peripherally circulating tumour cells, a variable fraction is able to express stem cell and adhesion properties and can be grown into tumour spheres, a property ascribed to cells capable of initiating tumours and metastases.

View Article: PubMed Central - PubMed

Affiliation: Transfusion Center Bayreuth D-95448, Germany.

ABSTRACT

Background: The detection of tumour cells circulating in the peripheral blood of patients with breast cancer is a sign that cells have been able to leave the primary tumour and survive in the circulation. However, in order to form metastases, they require additional properties such as the ability to adhere, self-renew, and grow. Here we present data that a variable fraction among the circulating tumour cells detected by the Maintrac(®) approach expresses mRNA of the stem cell gene NANOG and of the adhesion molecule vimentin and is capable of forming tumour spheres, a property ascribed to tumour-initiating cells (TICs).

Patients and methods: Between ten and 50 circulating epithelial antigen-positive cells detected by the Maintrac approach were selected randomly from each of 20 patients with breast cancer before and after surgery and were isolated using automated capillary aspiration and deposited individually onto slides for expression profiling. In addition, the circulating tumour cells were cultured without isolation among the white blood cells from 39 patients with breast cancer in different stages of disease using culture methods favouring growth of epithelial cells.

Results: Although no epithelial cell adhesion molecule (EpCAM)-positive cells expressing stem cell genes or the adhesion molecule vimentin was detected before surgery, 10%-20% of the cells were found to be positive for mRNA of these genes after surgery. Tumour spheres from circulating cells of 39 patients with different stages of breast cancer were grown without previous isolation in a fraction increasing with the aggressivity of the tumour.

Summary: Here we show that among the peripherally circulating tumour cells, a variable fraction is able to express stem cell and adhesion properties and can be grown into tumour spheres, a property ascribed to cells capable of initiating tumours and metastases.

No MeSH data available.


Related in: MedlinePlus