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Mammary carcinoma behavior is programmed in the precancer stem cell.

Damonte P, Hodgson JG, Chen JQ, Young LJ, Cardiff RD, Borowsky AD - Breast Cancer Res. (2008)

Bottom Line: No changes are associated with progression, although spontaneous focal amplifications and deletions were detected occasionally.Telomerase activity is increased in both the MINO and the derived tumors when compared with normal mouse mammary gland.Within the context of this model, these cells have programmed potential for latency and metastasis that does not appear to require sequential genetic 'hits' for transformation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, Center for Comparative Medicine, UC Davis, County Road 98 and Hutchison Drive, Davis, California 95616, USA.

ABSTRACT

Introduction: The 'MINO' (mammary intraepithelial neoplasia outgrowth) mouse model of ductal carcinoma in situ (DCIS) consists of six lines with distinct morphologic phenotypes and behavior, each meeting experimentally defined criteria for 'precancer'. Specifically, these lines grow orthotopically in cleared mammary fat pads and consistently progress to an invasive phenotype that is capable of ectopic growth. Transition to carcinoma has a consistent latency for each line, and three of the lines also exhibit pulmonary metastatic potential.

Methods: Gland cleared orthotopic transplanted precancer MINO tissues were analyzed by bacterial artifical chromosome and oligo array comparative genomic hybridization, microsatellite PCR, and telomerase repeat amplification assay. MINO cells were dissociated and cultured in three dimensional culture and transplanted in syngeneic gland cleared mammary fat pads.

Results: Comparative genomic hybridization shows that the precancer and invasive tumors are genetically stable, with low level changes including whole chromosome gains in some lines. No changes are associated with progression, although spontaneous focal amplifications and deletions were detected occasionally. Microsatellite analysis shows a low frequency of alterations that are predominantly permanent within a MINO line. Telomerase activity is increased in both the MINO and the derived tumors when compared with normal mouse mammary gland. Dissociation of the precancer lesion cells and three dimensional 'spheroid' culture of single cells reveals a bipotential for myoepithelial and luminal differentiation and the formation of unique three-dimensional 'MINOspheres'. These MINOspheres exhibit features that are intermediate between spheroids that are derived from normal and carcinoma cells. Transplantation of a single cell derived MINOsphere recapitulates the outgrowth of the precancer morphology and progression to carcinoma.

Conclusion: These data establish a precancer 'stem' cell that is capable of self-renewal and multilineage differentiation as the origin of invasive cancer. Within the context of this model, these cells have programmed potential for latency and metastasis that does not appear to require sequential genetic 'hits' for transformation.

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Immunohistochemistry profile of MINOsphere-derived outgrowth versus the original MINO tissue outgrowth. The outgrowth derived from a MINOsphere transplant (a, c, e, g, i, k) shows analogous expression compared with the MINO tissue outgrowth from the donor (b, d, f, h, j, m) for the following: luminal epithelial, CK8–18 (panels a and b); basal epithelial/myoepithelial, CK14 (panels c and d) and CK5 (panels g and h); myoepithelial/smooth muscle, SMA (panels e and f insets show higher magnification of SMA-positive cells within the epithelial layers); and proliferation/apoptosis, Ki67 (panels I and j)/caspase3 (panels k and l). Panels a to h are of identical magnification, with the 200 μm scale bar shown (panel h). Panels i to l are identical magnification with the 100 μm scale bar shown (panel l). CK, cytokeratin; MINO, mammary intraepithelial neoplasia outgrowth; SMA, smooth muscle actin.
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Figure 4: Immunohistochemistry profile of MINOsphere-derived outgrowth versus the original MINO tissue outgrowth. The outgrowth derived from a MINOsphere transplant (a, c, e, g, i, k) shows analogous expression compared with the MINO tissue outgrowth from the donor (b, d, f, h, j, m) for the following: luminal epithelial, CK8–18 (panels a and b); basal epithelial/myoepithelial, CK14 (panels c and d) and CK5 (panels g and h); myoepithelial/smooth muscle, SMA (panels e and f insets show higher magnification of SMA-positive cells within the epithelial layers); and proliferation/apoptosis, Ki67 (panels I and j)/caspase3 (panels k and l). Panels a to h are of identical magnification, with the 200 μm scale bar shown (panel h). Panels i to l are identical magnification with the 100 μm scale bar shown (panel l). CK, cytokeratin; MINO, mammary intraepithelial neoplasia outgrowth; SMA, smooth muscle actin.

Mentions: The phenotypes of MINOsphere derived outhgrowths matched the original (donor serial transplant) outgrowths in comparative pathology analysis. This comparison was also validated by immunohistochemistry. The immunohistochemistry analyses confirm the same patterns and distributions of expression of CK8–18, CK5, CK14, and SMA. Ki67 and activated caspase 3 immunohistochemistry also showed matching proliferation and apoptotic rates (Figure 4). Ki67 quantification in the proliferation zone revealed a comparable proliferation percentage in donor samples (64.1% ± 20%) and MINOsphere derived outhgrowths (62% ± 28%). As expected, some histologic pattern variation between different MINO lines was observed, and the variation matched patterns characteristic of each line. A sample comparison of the phenotype of derivative donor MINO line and the single cell transplant for MINO line 4 is illustrated in Figure 4. Additionally, second and third serial transplantations from the single cell MINOsphere-derived outgrowths retained their characteristic morphologies (Additional file 3).


Mammary carcinoma behavior is programmed in the precancer stem cell.

Damonte P, Hodgson JG, Chen JQ, Young LJ, Cardiff RD, Borowsky AD - Breast Cancer Res. (2008)

Immunohistochemistry profile of MINOsphere-derived outgrowth versus the original MINO tissue outgrowth. The outgrowth derived from a MINOsphere transplant (a, c, e, g, i, k) shows analogous expression compared with the MINO tissue outgrowth from the donor (b, d, f, h, j, m) for the following: luminal epithelial, CK8–18 (panels a and b); basal epithelial/myoepithelial, CK14 (panels c and d) and CK5 (panels g and h); myoepithelial/smooth muscle, SMA (panels e and f insets show higher magnification of SMA-positive cells within the epithelial layers); and proliferation/apoptosis, Ki67 (panels I and j)/caspase3 (panels k and l). Panels a to h are of identical magnification, with the 200 μm scale bar shown (panel h). Panels i to l are identical magnification with the 100 μm scale bar shown (panel l). CK, cytokeratin; MINO, mammary intraepithelial neoplasia outgrowth; SMA, smooth muscle actin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2481504&req=5

Figure 4: Immunohistochemistry profile of MINOsphere-derived outgrowth versus the original MINO tissue outgrowth. The outgrowth derived from a MINOsphere transplant (a, c, e, g, i, k) shows analogous expression compared with the MINO tissue outgrowth from the donor (b, d, f, h, j, m) for the following: luminal epithelial, CK8–18 (panels a and b); basal epithelial/myoepithelial, CK14 (panels c and d) and CK5 (panels g and h); myoepithelial/smooth muscle, SMA (panels e and f insets show higher magnification of SMA-positive cells within the epithelial layers); and proliferation/apoptosis, Ki67 (panels I and j)/caspase3 (panels k and l). Panels a to h are of identical magnification, with the 200 μm scale bar shown (panel h). Panels i to l are identical magnification with the 100 μm scale bar shown (panel l). CK, cytokeratin; MINO, mammary intraepithelial neoplasia outgrowth; SMA, smooth muscle actin.
Mentions: The phenotypes of MINOsphere derived outhgrowths matched the original (donor serial transplant) outgrowths in comparative pathology analysis. This comparison was also validated by immunohistochemistry. The immunohistochemistry analyses confirm the same patterns and distributions of expression of CK8–18, CK5, CK14, and SMA. Ki67 and activated caspase 3 immunohistochemistry also showed matching proliferation and apoptotic rates (Figure 4). Ki67 quantification in the proliferation zone revealed a comparable proliferation percentage in donor samples (64.1% ± 20%) and MINOsphere derived outhgrowths (62% ± 28%). As expected, some histologic pattern variation between different MINO lines was observed, and the variation matched patterns characteristic of each line. A sample comparison of the phenotype of derivative donor MINO line and the single cell transplant for MINO line 4 is illustrated in Figure 4. Additionally, second and third serial transplantations from the single cell MINOsphere-derived outgrowths retained their characteristic morphologies (Additional file 3).

Bottom Line: No changes are associated with progression, although spontaneous focal amplifications and deletions were detected occasionally.Telomerase activity is increased in both the MINO and the derived tumors when compared with normal mouse mammary gland.Within the context of this model, these cells have programmed potential for latency and metastasis that does not appear to require sequential genetic 'hits' for transformation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, Center for Comparative Medicine, UC Davis, County Road 98 and Hutchison Drive, Davis, California 95616, USA.

ABSTRACT

Introduction: The 'MINO' (mammary intraepithelial neoplasia outgrowth) mouse model of ductal carcinoma in situ (DCIS) consists of six lines with distinct morphologic phenotypes and behavior, each meeting experimentally defined criteria for 'precancer'. Specifically, these lines grow orthotopically in cleared mammary fat pads and consistently progress to an invasive phenotype that is capable of ectopic growth. Transition to carcinoma has a consistent latency for each line, and three of the lines also exhibit pulmonary metastatic potential.

Methods: Gland cleared orthotopic transplanted precancer MINO tissues were analyzed by bacterial artifical chromosome and oligo array comparative genomic hybridization, microsatellite PCR, and telomerase repeat amplification assay. MINO cells were dissociated and cultured in three dimensional culture and transplanted in syngeneic gland cleared mammary fat pads.

Results: Comparative genomic hybridization shows that the precancer and invasive tumors are genetically stable, with low level changes including whole chromosome gains in some lines. No changes are associated with progression, although spontaneous focal amplifications and deletions were detected occasionally. Microsatellite analysis shows a low frequency of alterations that are predominantly permanent within a MINO line. Telomerase activity is increased in both the MINO and the derived tumors when compared with normal mouse mammary gland. Dissociation of the precancer lesion cells and three dimensional 'spheroid' culture of single cells reveals a bipotential for myoepithelial and luminal differentiation and the formation of unique three-dimensional 'MINOspheres'. These MINOspheres exhibit features that are intermediate between spheroids that are derived from normal and carcinoma cells. Transplantation of a single cell derived MINOsphere recapitulates the outgrowth of the precancer morphology and progression to carcinoma.

Conclusion: These data establish a precancer 'stem' cell that is capable of self-renewal and multilineage differentiation as the origin of invasive cancer. Within the context of this model, these cells have programmed potential for latency and metastasis that does not appear to require sequential genetic 'hits' for transformation.

Show MeSH
Related in: MedlinePlus