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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

Breast tumors in TNBC patients show higher expression of centrosomal proteins and aggressiveness markers(A) Micrographs showing immunohistchemical staining for Aurora-A, Plk4 (centrosomal amplification markers) and vimentin (breast cancer metastasis marker) in normal and cancer tissue from representative grade-matched TNBC (2170) and non-TNBC (6456) patients. (B) Box whisker graph showing significantly higher expression of vimentin in TNBC samples when compared to grade-matched non-TNBC samples. (C) Immunoblots for normal, TNBC (2170) and non-TNBC (6456) tissue samples showing Aurora-A, Cyclin E and vimentin expression. (D) Progression free survival (PFS) plot for the patients with higher (> 20%) CA (red) and lower (< 20%) CA (blue). (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 4: Breast tumors in TNBC patients show higher expression of centrosomal proteins and aggressiveness markers(A) Micrographs showing immunohistchemical staining for Aurora-A, Plk4 (centrosomal amplification markers) and vimentin (breast cancer metastasis marker) in normal and cancer tissue from representative grade-matched TNBC (2170) and non-TNBC (6456) patients. (B) Box whisker graph showing significantly higher expression of vimentin in TNBC samples when compared to grade-matched non-TNBC samples. (C) Immunoblots for normal, TNBC (2170) and non-TNBC (6456) tissue samples showing Aurora-A, Cyclin E and vimentin expression. (D) Progression free survival (PFS) plot for the patients with higher (> 20%) CA (red) and lower (< 20%) CA (blue). (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 evaluated expression levels of molecules implicated in CA by immunohistochemical staining of paraffin-embedded tumor and uninvolved adjacent tissue in 20 pairs of grade-matched (Grade II) TNBC and non-TNBC samples. Immunohistochemical data strongly suggested overexpression of well-established centrosome amplification markers (Aurora-A and Plk4) in TNBC compared to non-TNBC samples (Fig. 4A). The same samples were also immunostained for vimentin, a breast cancer aggressiveness marker. As expected, TNBC samples exhibited significantly higher expression of vimentin (Fig. 4B). Representative images of two sets of grade-matched breast tumor samples (one TNBC, one non-TNBC) exhibiting different expression levels of centrosomal proteins, show that TNBC samples exhibited higher expression levels of these proteins than non-TNBC samples and normal tissue (Fig. 4A). Statistical analysis of 20 pairs of grade-matched TNBC and non-TNBC samples revealed a strong positive correlation between Plk4 and vimentin (r = 0.58, p < 0.02). Immunoblots shown in Fig. 4C also support our observation that centrosomal markers (Aurora-A, Plk4 and cyclin E) exhibit similar expression patterns and trends as metastasis markers (vimentin) in breast tumor samples. Furthermore, we evaluated the relationship of higher centrosome aberrations (as assessed by %CA in Fig. 2Bi) with progression-free survival (PFS) in breast cancer patients. PFS was calculated as the number of days from diagnosis to the first local recurrence or metastasis (if one occurred), or the last follow-up if the patient did not progress. Irrespective of receptor status (n = 120), patients with higher centrosome aberrations (> 20%) had lower PFS (p < 0.08) than patients with lower centrosome aberrations (< 20%) (Fig. 4D). Interestingly, a majority (~72%) of high CA (high-risk) group were TNBC while low CA (low-risk) group were largely non-TNBC (~60%).


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)

Breast tumors in TNBC patients show higher expression of centrosomal proteins and aggressiveness markers(A) Micrographs showing immunohistchemical staining for Aurora-A, Plk4 (centrosomal amplification markers) and vimentin (breast cancer metastasis marker) in normal and cancer tissue from representative grade-matched TNBC (2170) and non-TNBC (6456) patients. (B) Box whisker graph showing significantly higher expression of vimentin in TNBC samples when compared to grade-matched non-TNBC samples. (C) Immunoblots for normal, TNBC (2170) and non-TNBC (6456) tissue samples showing Aurora-A, Cyclin E and vimentin expression. (D) Progression free survival (PFS) plot for the patients with higher (> 20%) CA (red) and lower (< 20%) CA (blue). (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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Related In: Results  -  Collection

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Show All Figures
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Figure 4: Breast tumors in TNBC patients show higher expression of centrosomal proteins and aggressiveness markers(A) Micrographs showing immunohistchemical staining for Aurora-A, Plk4 (centrosomal amplification markers) and vimentin (breast cancer metastasis marker) in normal and cancer tissue from representative grade-matched TNBC (2170) and non-TNBC (6456) patients. (B) Box whisker graph showing significantly higher expression of vimentin in TNBC samples when compared to grade-matched non-TNBC samples. (C) Immunoblots for normal, TNBC (2170) and non-TNBC (6456) tissue samples showing Aurora-A, Cyclin E and vimentin expression. (D) Progression free survival (PFS) plot for the patients with higher (> 20%) CA (red) and lower (< 20%) CA (blue). (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 evaluated expression levels of molecules implicated in CA by immunohistochemical staining of paraffin-embedded tumor and uninvolved adjacent tissue in 20 pairs of grade-matched (Grade II) TNBC and non-TNBC samples. Immunohistochemical data strongly suggested overexpression of well-established centrosome amplification markers (Aurora-A and Plk4) in TNBC compared to non-TNBC samples (Fig. 4A). The same samples were also immunostained for vimentin, a breast cancer aggressiveness marker. As expected, TNBC samples exhibited significantly higher expression of vimentin (Fig. 4B). Representative images of two sets of grade-matched breast tumor samples (one TNBC, one non-TNBC) exhibiting different expression levels of centrosomal proteins, show that TNBC samples exhibited higher expression levels of these proteins than non-TNBC samples and normal tissue (Fig. 4A). Statistical analysis of 20 pairs of grade-matched TNBC and non-TNBC samples revealed a strong positive correlation between Plk4 and vimentin (r = 0.58, p < 0.02). Immunoblots shown in Fig. 4C also support our observation that centrosomal markers (Aurora-A, Plk4 and cyclin E) exhibit similar expression patterns and trends as metastasis markers (vimentin) in breast tumor samples. Furthermore, we evaluated the relationship of higher centrosome aberrations (as assessed by %CA in Fig. 2Bi) with progression-free survival (PFS) in breast cancer patients. PFS was calculated as the number of days from diagnosis to the first local recurrence or metastasis (if one occurred), or the last follow-up if the patient did not progress. Irrespective of receptor status (n = 120), patients with higher centrosome aberrations (> 20%) had lower PFS (p < 0.08) than patients with lower centrosome aberrations (< 20%) (Fig. 4D). Interestingly, a majority (~72%) of high CA (high-risk) group were TNBC while low CA (low-risk) group were largely non-TNBC (~60%).

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