Human mammary cancer progression model recapitulates methylation events associated with breast premalignancy

Nancy Dumont, Yongping G. Crawford, Mahvash Sigaroudinia, Shefali S. Nagrani, Matthew B. Wilson, Gertrude C. Buehring, Gulisa Turashvili, Samuel Aparicio, Mona L. Gauthier, Colleen A. Fordyce, Kimberly McDermott, Thea D. Tlsty

Research output: Contribution to journalArticle

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Abstract

Introduction: We have previously identified a rare subpopulation of variant human mammary epithelial cells (vHMEC) with repressed p16INK4A that exist in disease-free women yet display premalignant properties, suggesting that they have engaged the process of malignant transformation. In order to gain insight into the molecular alterations required for vHMEC to progress to malignancy, and to characterize the epigenetic events associated with early progression, we examined the effect of oncogenic stress on the behavior of these cells.Methods: HMEC that express p16INK4A and vHMEC that do not, were transduced with constitutively active Ha-rasV12 and subsequently exposed to serum to determine whether signals from the cellular microenvironment could cooperate with ras to promote the malignant transformation of vHMEC. Epigenetic alterations were assessed using methylation-specific polymerase chain reaction (PCR).Results: vHMEC expressing Ha-rasV12 (vHMEC-ras) bypassed the classic proliferative arrest that has been previously documented in normal fibroblasts following oncogenic stress, and that we also observe here in normal HMEC. Moreover, vHMEC-ras cells exhibited many additional alterations that are observed during progression to malignancy such as the generation of chromosomal abnormalities, upregulation of telomerase activity, immortalization following exposure to serum, and anchorage-independent growth, but they did not form tumors following orthotopic injection in vivo. Associated with their early progression to malignancy was an increase in the number of genes methylated, two of which (RASSF1A and SFRP1) were also methylated in other immortalized mammary cell lines as well as in breast cancer cells and tissues.Conclusions: We have characterized a mammary progression model that recapitulates molecular and methylation alterations observed in many breast cancers. Our data suggest that concomitant methylation of RASSF1A and SFRP1 marks an early event in mammary transformation and may thus have prognostic potential.

Original languageEnglish (US)
Article numberR87
JournalBreast Cancer Research
Volume11
Issue number6
DOIs
StatePublished - Dec 8 2009

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Methylation
Breast
Breast Neoplasms
Epithelial Cells
Epigenomics
Neoplasms
Cellular Microenvironment
Molecular Models
Telomerase
Serum
Chromosome Aberrations
Up-Regulation
Fibroblasts
Cell Line
Polymerase Chain Reaction
Injections
Growth
Genes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Medicine(all)

Cite this

Dumont, N., Crawford, Y. G., Sigaroudinia, M., Nagrani, S. S., Wilson, M. B., Buehring, G. C., ... Tlsty, T. D. (2009). Human mammary cancer progression model recapitulates methylation events associated with breast premalignancy. Breast Cancer Research, 11(6), [R87]. https://doi.org/10.1186/bcr2457

Human mammary cancer progression model recapitulates methylation events associated with breast premalignancy. / Dumont, Nancy; Crawford, Yongping G.; Sigaroudinia, Mahvash; Nagrani, Shefali S.; Wilson, Matthew B.; Buehring, Gertrude C.; Turashvili, Gulisa; Aparicio, Samuel; Gauthier, Mona L.; Fordyce, Colleen A.; McDermott, Kimberly; Tlsty, Thea D.

In: Breast Cancer Research, Vol. 11, No. 6, R87, 08.12.2009.

Research output: Contribution to journalArticle

Dumont, N, Crawford, YG, Sigaroudinia, M, Nagrani, SS, Wilson, MB, Buehring, GC, Turashvili, G, Aparicio, S, Gauthier, ML, Fordyce, CA, McDermott, K & Tlsty, TD 2009, 'Human mammary cancer progression model recapitulates methylation events associated with breast premalignancy', Breast Cancer Research, vol. 11, no. 6, R87. https://doi.org/10.1186/bcr2457
Dumont N, Crawford YG, Sigaroudinia M, Nagrani SS, Wilson MB, Buehring GC et al. Human mammary cancer progression model recapitulates methylation events associated with breast premalignancy. Breast Cancer Research. 2009 Dec 8;11(6). R87. https://doi.org/10.1186/bcr2457
Dumont, Nancy ; Crawford, Yongping G. ; Sigaroudinia, Mahvash ; Nagrani, Shefali S. ; Wilson, Matthew B. ; Buehring, Gertrude C. ; Turashvili, Gulisa ; Aparicio, Samuel ; Gauthier, Mona L. ; Fordyce, Colleen A. ; McDermott, Kimberly ; Tlsty, Thea D. / Human mammary cancer progression model recapitulates methylation events associated with breast premalignancy. In: Breast Cancer Research. 2009 ; Vol. 11, No. 6.
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abstract = "Introduction: We have previously identified a rare subpopulation of variant human mammary epithelial cells (vHMEC) with repressed p16INK4A that exist in disease-free women yet display premalignant properties, suggesting that they have engaged the process of malignant transformation. In order to gain insight into the molecular alterations required for vHMEC to progress to malignancy, and to characterize the epigenetic events associated with early progression, we examined the effect of oncogenic stress on the behavior of these cells.Methods: HMEC that express p16INK4A and vHMEC that do not, were transduced with constitutively active Ha-rasV12 and subsequently exposed to serum to determine whether signals from the cellular microenvironment could cooperate with ras to promote the malignant transformation of vHMEC. Epigenetic alterations were assessed using methylation-specific polymerase chain reaction (PCR).Results: vHMEC expressing Ha-rasV12 (vHMEC-ras) bypassed the classic proliferative arrest that has been previously documented in normal fibroblasts following oncogenic stress, and that we also observe here in normal HMEC. Moreover, vHMEC-ras cells exhibited many additional alterations that are observed during progression to malignancy such as the generation of chromosomal abnormalities, upregulation of telomerase activity, immortalization following exposure to serum, and anchorage-independent growth, but they did not form tumors following orthotopic injection in vivo. Associated with their early progression to malignancy was an increase in the number of genes methylated, two of which (RASSF1A and SFRP1) were also methylated in other immortalized mammary cell lines as well as in breast cancer cells and tissues.Conclusions: We have characterized a mammary progression model that recapitulates molecular and methylation alterations observed in many breast cancers. Our data suggest that concomitant methylation of RASSF1A and SFRP1 marks an early event in mammary transformation and may thus have prognostic potential.",
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AU - Crawford, Yongping G.

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AU - Wilson, Matthew B.

AU - Buehring, Gertrude C.

AU - Turashvili, Gulisa

AU - Aparicio, Samuel

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N2 - Introduction: We have previously identified a rare subpopulation of variant human mammary epithelial cells (vHMEC) with repressed p16INK4A that exist in disease-free women yet display premalignant properties, suggesting that they have engaged the process of malignant transformation. In order to gain insight into the molecular alterations required for vHMEC to progress to malignancy, and to characterize the epigenetic events associated with early progression, we examined the effect of oncogenic stress on the behavior of these cells.Methods: HMEC that express p16INK4A and vHMEC that do not, were transduced with constitutively active Ha-rasV12 and subsequently exposed to serum to determine whether signals from the cellular microenvironment could cooperate with ras to promote the malignant transformation of vHMEC. Epigenetic alterations were assessed using methylation-specific polymerase chain reaction (PCR).Results: vHMEC expressing Ha-rasV12 (vHMEC-ras) bypassed the classic proliferative arrest that has been previously documented in normal fibroblasts following oncogenic stress, and that we also observe here in normal HMEC. Moreover, vHMEC-ras cells exhibited many additional alterations that are observed during progression to malignancy such as the generation of chromosomal abnormalities, upregulation of telomerase activity, immortalization following exposure to serum, and anchorage-independent growth, but they did not form tumors following orthotopic injection in vivo. Associated with their early progression to malignancy was an increase in the number of genes methylated, two of which (RASSF1A and SFRP1) were also methylated in other immortalized mammary cell lines as well as in breast cancer cells and tissues.Conclusions: We have characterized a mammary progression model that recapitulates molecular and methylation alterations observed in many breast cancers. Our data suggest that concomitant methylation of RASSF1A and SFRP1 marks an early event in mammary transformation and may thus have prognostic potential.

AB - Introduction: We have previously identified a rare subpopulation of variant human mammary epithelial cells (vHMEC) with repressed p16INK4A that exist in disease-free women yet display premalignant properties, suggesting that they have engaged the process of malignant transformation. In order to gain insight into the molecular alterations required for vHMEC to progress to malignancy, and to characterize the epigenetic events associated with early progression, we examined the effect of oncogenic stress on the behavior of these cells.Methods: HMEC that express p16INK4A and vHMEC that do not, were transduced with constitutively active Ha-rasV12 and subsequently exposed to serum to determine whether signals from the cellular microenvironment could cooperate with ras to promote the malignant transformation of vHMEC. Epigenetic alterations were assessed using methylation-specific polymerase chain reaction (PCR).Results: vHMEC expressing Ha-rasV12 (vHMEC-ras) bypassed the classic proliferative arrest that has been previously documented in normal fibroblasts following oncogenic stress, and that we also observe here in normal HMEC. Moreover, vHMEC-ras cells exhibited many additional alterations that are observed during progression to malignancy such as the generation of chromosomal abnormalities, upregulation of telomerase activity, immortalization following exposure to serum, and anchorage-independent growth, but they did not form tumors following orthotopic injection in vivo. Associated with their early progression to malignancy was an increase in the number of genes methylated, two of which (RASSF1A and SFRP1) were also methylated in other immortalized mammary cell lines as well as in breast cancer cells and tissues.Conclusions: We have characterized a mammary progression model that recapitulates molecular and methylation alterations observed in many breast cancers. Our data suggest that concomitant methylation of RASSF1A and SFRP1 marks an early event in mammary transformation and may thus have prognostic potential.

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