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Rampant centrosome amplification underlies more aggressive disease course of triple negative breast cancers.

Pannu V, Mittal K, Cantuaria G, Reid MD, Li X, Donthamsetty S, McBride M, Klimov S, Osan R, Gupta MV, Rida PC, Aneja R - Oncotarget (2015)

Bottom Line: Our data establish differences in incidence and severity of CA between TNBC and non-TNBC cell lines and clinical specimens.We found strong correlation between CA and aggressiveness markers associated with metastasis in 20 pairs of grade-matched TNBC and non-TNBC specimens (p < 0.02).Time-lapse imaging of MDA-MB-231 cells harboring amplified centrosomes demonstrated enhanced migratory ability.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Georgia State University, Atlanta, GA 30303, USA.

ABSTRACT
Centrosome amplification (CA), a cell-biological trait, characterizes pre-neoplastic and pre-invasive lesions and is associated with tumor aggressiveness. Recent studies suggest that CA leads to malignant transformation and promotes invasion in mammary epithelial cells. Triple negative breast cancer (TNBC), a histologically-aggressive subtype shows high recurrence, metastases, and mortality rates. Since TNBC and non-TNBC follow variable kinetics of metastatic progression, they constitute a novel test bed to explore if severity and nature of CA can distinguish them apart. We quantitatively assessed structural and numerical centrosomal aberrations for each patient sample in a large-cohort of grade-matched TNBC (n = 30) and non-TNBC (n = 98) cases employing multi-color confocal imaging. Our data establish differences in incidence and severity of CA between TNBC and non-TNBC cell lines and clinical specimens. We found strong correlation between CA and aggressiveness markers associated with metastasis in 20 pairs of grade-matched TNBC and non-TNBC specimens (p < 0.02). Time-lapse imaging of MDA-MB-231 cells harboring amplified centrosomes demonstrated enhanced migratory ability. Our study bridges a vital knowledge gap by pinpointing that CA underlies breast cancer aggressiveness. This previously unrecognized organellar inequality at the centrosome level may allow early-risk prediction and explain higher tumor aggressiveness and mortality rates in TNBC patients.

No MeSH data available.


Related in: MedlinePlus

TNBC tumors and cell lines show higher expression of centrosomal markers(A) Immunoblots for 5 paired breast tumor (T) and normal adjacent (N) tissues from grade-matched TNBC and non-TNBC patients showing expression levels of centrosomal markers. (B) Immunoblots of centrosomal markers in MCF-7 (non-TNBC), MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Ci) Immunofluorescence micrographs showing MCF-10A cells (near-normal), MCF-7 cells (non-TNBC) and MDA-MB-468 and MDA-MB-231 cells (TNBC) in interphase and mitotic state, stained for γ-tubulin (green), α-tubulin (red) and DAPI (blue). (Cii) Bar graph quantitation of percent cells depicting multipolar mitosis and centrosomal amplification in interphase in MCF-7 (non-TNBC) and MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Only Grade II TNBC and non-TNBC samples were matched because of limited n numbers for Grade I TNBC and Grade III non-TNBC samples in our dataset.)
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Figure 3: TNBC tumors and cell lines show higher expression of centrosomal markers(A) Immunoblots for 5 paired breast tumor (T) and normal adjacent (N) tissues from grade-matched TNBC and non-TNBC patients showing expression levels of centrosomal markers. (B) Immunoblots of centrosomal markers in MCF-7 (non-TNBC), MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Ci) Immunofluorescence micrographs showing MCF-10A cells (near-normal), MCF-7 cells (non-TNBC) and MDA-MB-468 and MDA-MB-231 cells (TNBC) in interphase and mitotic state, stained for γ-tubulin (green), α-tubulin (red) and DAPI (blue). (Cii) Bar graph quantitation of percent cells depicting multipolar mitosis and centrosomal amplification in interphase in MCF-7 (non-TNBC) and MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Only Grade II TNBC and non-TNBC samples were matched because of limited n numbers for Grade I TNBC and Grade III non-TNBC samples in our dataset.)

Mentions: We next compared the expression levels of centrosomal proteins (centrin-2 and γ-tubulin) in fresh-frozen tumors and uninvolved adjacent tissue from 20 pairs of grade-matched TNBC and non-TNBC patients. Immunoblotting of tissue lysates showed higher expression of two centrosomal proteins (centrin-2 and γ-tubulin) in TNBC than in non-TNBC patients (Fig. 3A). In addition, breast cancer cell lines derived from TNBC patients (MDA-MB-468, MDA-MB-231) showed (a) higher incidence and severity of centrosome amplification (Fig. 3Ci, ii) (b) elevated expression of centrosomal (pericentrin, centrin-2, γ-tubulin) and centrosome amplification markers (Plk4 and cyclin E), compared to non-TNBC-derived (MCF-7) lines (Fig. 3B). Intriguingly, the extent of amplification was considerably lower in cell lines (5–30%) compared to patient tissue samples (15–80%), irrespective of the receptor status. This observation highlights a previously unrecognized discordance between human tumor tissues and established cell lines. This also underscores the limitations of in vitro cell lines as a model system for establishing and testing centrosome-targeted therapies.


Rampant centrosome amplification underlies more aggressive disease course of triple negative breast cancers.

Pannu V, Mittal K, Cantuaria G, Reid MD, Li X, Donthamsetty S, McBride M, Klimov S, Osan R, Gupta MV, Rida PC, Aneja R - Oncotarget (2015)

TNBC tumors and cell lines show higher expression of centrosomal markers(A) Immunoblots for 5 paired breast tumor (T) and normal adjacent (N) tissues from grade-matched TNBC and non-TNBC patients showing expression levels of centrosomal markers. (B) Immunoblots of centrosomal markers in MCF-7 (non-TNBC), MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Ci) Immunofluorescence micrographs showing MCF-10A cells (near-normal), MCF-7 cells (non-TNBC) and MDA-MB-468 and MDA-MB-231 cells (TNBC) in interphase and mitotic state, stained for γ-tubulin (green), α-tubulin (red) and DAPI (blue). (Cii) Bar graph quantitation of percent cells depicting multipolar mitosis and centrosomal amplification in interphase in MCF-7 (non-TNBC) and MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Only Grade II TNBC and non-TNBC samples were matched because of limited n numbers for Grade I TNBC and Grade III non-TNBC samples in our dataset.)
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Figure 3: TNBC tumors and cell lines show higher expression of centrosomal markers(A) Immunoblots for 5 paired breast tumor (T) and normal adjacent (N) tissues from grade-matched TNBC and non-TNBC patients showing expression levels of centrosomal markers. (B) Immunoblots of centrosomal markers in MCF-7 (non-TNBC), MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Ci) Immunofluorescence micrographs showing MCF-10A cells (near-normal), MCF-7 cells (non-TNBC) and MDA-MB-468 and MDA-MB-231 cells (TNBC) in interphase and mitotic state, stained for γ-tubulin (green), α-tubulin (red) and DAPI (blue). (Cii) Bar graph quantitation of percent cells depicting multipolar mitosis and centrosomal amplification in interphase in MCF-7 (non-TNBC) and MDA-MB-468 and MDA-MB-231 (TNBC) cell lines. (Only Grade II TNBC and non-TNBC samples were matched because of limited n numbers for Grade I TNBC and Grade III non-TNBC samples in our dataset.)
Mentions: We next compared the expression levels of centrosomal proteins (centrin-2 and γ-tubulin) in fresh-frozen tumors and uninvolved adjacent tissue from 20 pairs of grade-matched TNBC and non-TNBC patients. Immunoblotting of tissue lysates showed higher expression of two centrosomal proteins (centrin-2 and γ-tubulin) in TNBC than in non-TNBC patients (Fig. 3A). In addition, breast cancer cell lines derived from TNBC patients (MDA-MB-468, MDA-MB-231) showed (a) higher incidence and severity of centrosome amplification (Fig. 3Ci, ii) (b) elevated expression of centrosomal (pericentrin, centrin-2, γ-tubulin) and centrosome amplification markers (Plk4 and cyclin E), compared to non-TNBC-derived (MCF-7) lines (Fig. 3B). Intriguingly, the extent of amplification was considerably lower in cell lines (5–30%) compared to patient tissue samples (15–80%), irrespective of the receptor status. This observation highlights a previously unrecognized discordance between human tumor tissues and established cell lines. This also underscores the limitations of in vitro cell lines as a model system for establishing and testing centrosome-targeted therapies.

Bottom Line: Our data establish differences in incidence and severity of CA between TNBC and non-TNBC cell lines and clinical specimens.We found strong correlation between CA and aggressiveness markers associated with metastasis in 20 pairs of grade-matched TNBC and non-TNBC specimens (p < 0.02).Time-lapse imaging of MDA-MB-231 cells harboring amplified centrosomes demonstrated enhanced migratory ability.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Georgia State University, Atlanta, GA 30303, USA.

ABSTRACT
Centrosome amplification (CA), a cell-biological trait, characterizes pre-neoplastic and pre-invasive lesions and is associated with tumor aggressiveness. Recent studies suggest that CA leads to malignant transformation and promotes invasion in mammary epithelial cells. Triple negative breast cancer (TNBC), a histologically-aggressive subtype shows high recurrence, metastases, and mortality rates. Since TNBC and non-TNBC follow variable kinetics of metastatic progression, they constitute a novel test bed to explore if severity and nature of CA can distinguish them apart. We quantitatively assessed structural and numerical centrosomal aberrations for each patient sample in a large-cohort of grade-matched TNBC (n = 30) and non-TNBC (n = 98) cases employing multi-color confocal imaging. Our data establish differences in incidence and severity of CA between TNBC and non-TNBC cell lines and clinical specimens. We found strong correlation between CA and aggressiveness markers associated with metastasis in 20 pairs of grade-matched TNBC and non-TNBC specimens (p < 0.02). Time-lapse imaging of MDA-MB-231 cells harboring amplified centrosomes demonstrated enhanced migratory ability. Our study bridges a vital knowledge gap by pinpointing that CA underlies breast cancer aggressiveness. This previously unrecognized organellar inequality at the centrosome level may allow early-risk prediction and explain higher tumor aggressiveness and mortality rates in TNBC patients.

No MeSH data available.


Related in: MedlinePlus