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Purified malignant mammary epithelial cells maintain hormone responsiveness in culture.

Kothari MS, Ali S, Buluwela L, Livni N, Shousha S, Sinnett HD, Vashisht R, Thorpe P, Van Noorden S, Coombes RC, Slade MJ - Br. J. Cancer (2003)

Bottom Line: While the malignant cells can be maintained in culture they do not proliferate in contrast to purified breast epithelial cells from reduction mammoplasties.Four out of the seven ERalpha-positive tumours showed a significant reduction in cell numbers after tamoxifen treatment compared to oestradiol, ERalpha negative tumours failed to show a response.We conclude that (a) it is possible to purify and maintain breast cancer cells for a sufficient period to permit functional studies and (b) ERalpha is retained in culture facilitating the use of these cells in studies of the mechanism of endocrine response and resistance in vitro.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Cell Biology, Imperial College, London, UK.

ABSTRACT
Currently, the therapy for breast cancer is determined by immunohistochemical staining of the primary tumour for oestrogen receptor alpha (ERalpha). However, a proportion of ERalpha-positive patients fail to respond to tamoxifen and a proportion of ERalpha-negative patients show response. Here, we describe a novel procedure for the purification of malignant breast epithelial cells in an attempt to identify these patients at an early stage. Using this procedure, we are able to purify malignant cells to >90% purity as determined by immunohistochemical staining, cytology and fluorescent in situ hybridisation (FISH). While the malignant cells can be maintained in culture they do not proliferate in contrast to purified breast epithelial cells from reduction mammoplasties. Moreover, ERalpha and progesterone receptor (PR) expression is maintained in malignant cells, whereas normal epithelial cells rapidly lose ERalpha and PR. Functional studies were performed on the separated malignant cells in terms of their response to oestradiol and tamoxifen. Four out of the seven ERalpha-positive tumours showed a significant reduction in cell numbers after tamoxifen treatment compared to oestradiol, ERalpha negative tumours failed to show a response. We conclude that (a) it is possible to purify and maintain breast cancer cells for a sufficient period to permit functional studies and (b) ERalpha is retained in culture facilitating the use of these cells in studies of the mechanism of endocrine response and resistance in vitro.

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Graph showing the viability of primary normal and malignant mammary epithelial cells over 9 days as determined by trypan blue staining and cell counting. The experiment started 3 days after the cells were seeded in order to allow for cell adherence. The adherent normal epithelial cells (▪) proliferate and hence the increase in numbers after the initial reduction between days 0 and 3. Approximately 60% of the malignant epithelial cells were adherent after 3 days and this fell to approximately 45% 9 days after seeding (▵). The number of floating dead malignant cells (□) and the floating live malignant cells (•) was low throughout the experiment.
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fig3: Graph showing the viability of primary normal and malignant mammary epithelial cells over 9 days as determined by trypan blue staining and cell counting. The experiment started 3 days after the cells were seeded in order to allow for cell adherence. The adherent normal epithelial cells (▪) proliferate and hence the increase in numbers after the initial reduction between days 0 and 3. Approximately 60% of the malignant epithelial cells were adherent after 3 days and this fell to approximately 45% 9 days after seeding (▵). The number of floating dead malignant cells (□) and the floating live malignant cells (•) was low throughout the experiment.

Mentions: We examined the proportion of cells that retained viability and adherence in a subset of samples (n=3). Figure 3Figure 3


Purified malignant mammary epithelial cells maintain hormone responsiveness in culture.

Kothari MS, Ali S, Buluwela L, Livni N, Shousha S, Sinnett HD, Vashisht R, Thorpe P, Van Noorden S, Coombes RC, Slade MJ - Br. J. Cancer (2003)

Graph showing the viability of primary normal and malignant mammary epithelial cells over 9 days as determined by trypan blue staining and cell counting. The experiment started 3 days after the cells were seeded in order to allow for cell adherence. The adherent normal epithelial cells (▪) proliferate and hence the increase in numbers after the initial reduction between days 0 and 3. Approximately 60% of the malignant epithelial cells were adherent after 3 days and this fell to approximately 45% 9 days after seeding (▵). The number of floating dead malignant cells (□) and the floating live malignant cells (•) was low throughout the experiment.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376379&req=5

fig3: Graph showing the viability of primary normal and malignant mammary epithelial cells over 9 days as determined by trypan blue staining and cell counting. The experiment started 3 days after the cells were seeded in order to allow for cell adherence. The adherent normal epithelial cells (▪) proliferate and hence the increase in numbers after the initial reduction between days 0 and 3. Approximately 60% of the malignant epithelial cells were adherent after 3 days and this fell to approximately 45% 9 days after seeding (▵). The number of floating dead malignant cells (□) and the floating live malignant cells (•) was low throughout the experiment.
Mentions: We examined the proportion of cells that retained viability and adherence in a subset of samples (n=3). Figure 3Figure 3

Bottom Line: While the malignant cells can be maintained in culture they do not proliferate in contrast to purified breast epithelial cells from reduction mammoplasties.Four out of the seven ERalpha-positive tumours showed a significant reduction in cell numbers after tamoxifen treatment compared to oestradiol, ERalpha negative tumours failed to show a response.We conclude that (a) it is possible to purify and maintain breast cancer cells for a sufficient period to permit functional studies and (b) ERalpha is retained in culture facilitating the use of these cells in studies of the mechanism of endocrine response and resistance in vitro.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Cell Biology, Imperial College, London, UK.

ABSTRACT
Currently, the therapy for breast cancer is determined by immunohistochemical staining of the primary tumour for oestrogen receptor alpha (ERalpha). However, a proportion of ERalpha-positive patients fail to respond to tamoxifen and a proportion of ERalpha-negative patients show response. Here, we describe a novel procedure for the purification of malignant breast epithelial cells in an attempt to identify these patients at an early stage. Using this procedure, we are able to purify malignant cells to >90% purity as determined by immunohistochemical staining, cytology and fluorescent in situ hybridisation (FISH). While the malignant cells can be maintained in culture they do not proliferate in contrast to purified breast epithelial cells from reduction mammoplasties. Moreover, ERalpha and progesterone receptor (PR) expression is maintained in malignant cells, whereas normal epithelial cells rapidly lose ERalpha and PR. Functional studies were performed on the separated malignant cells in terms of their response to oestradiol and tamoxifen. Four out of the seven ERalpha-positive tumours showed a significant reduction in cell numbers after tamoxifen treatment compared to oestradiol, ERalpha negative tumours failed to show a response. We conclude that (a) it is possible to purify and maintain breast cancer cells for a sufficient period to permit functional studies and (b) ERalpha is retained in culture facilitating the use of these cells in studies of the mechanism of endocrine response and resistance in vitro.

Show MeSH
Related in: MedlinePlus