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NuMA overexpression in epithelial ovarian cancer.

Brüning-Richardson A, Bond J, Alsiary R, Richardson J, Cairns DA, McCormac L, Hutson R, Burns PA, Wilkinson N, Hall GD, Morrison EE, Bell SM - PLoS ONE (2012)

Bottom Line: IHC revealed low to weak NuMA expression in normal tissues.NuMA expression decreased in late disease stage 4 endometrioid EOCs.IF of primary cultures revealed that high NuMA levels at mitotic spindle poles were significantly associated with a decreased proportion of cells in cytokinesis (p = 0.05), increased binucleation (p = 0.021) and multinucleation (p = 0.007), and aneuploidy (p = 0.008).NuMA is highly expressed in EOC tumours and high NuMA levels correlate with increases in mitotic defects and aneuploidy in primary cultures.

View Article: PubMed Central - PubMed

Affiliation: Section of Ophthalmology and Neuroscience, Leeds Institute of Molecular Medicine, St James's University Hospital, Leeds, United Kingdom. bgyar@leeds.ac.uk

ABSTRACT
Highly aneuploid tumours are common in epithelial ovarian cancers (EOC). We investigated whether NuMA expression was associated with this phenomenon.NuMA protein levels in normal and tumour tissues, ovarian cell lines and primary cultures of malignant cells derived from ovarian ascitic fluids were analysed by Affymetrix microarray analysis, immunoblotting, immunohistochemistry (IHC) and immunofluorescence (IF), with results correlated to associated clinical data. Aneuploidy status in primary cultures was determined by FACS analysis.Affymetrix microarray data indicated that NuMA was overexpressed in tumour tissue, primary cultures and cell lines compared to normal ovarian tissue. IHC revealed low to weak NuMA expression in normal tissues. Expression was upregulated in tumours, with a significant association with disease stage in mucinous EOC subtypes (p = 0.009), lymph node involvement (p = 0.03) and patient age (p = 0.04). Additional discontinuous data analysis revealed that high NuMA levels in tumours decreased with grade (p = 0.02) but increased with disease stage (p = 0.04) in serous EOC. NuMA expression decreased in late disease stage 4 endometrioid EOCs. High NuMA levels decreased with increased tumour invasion in all subtypes (p = 0.03). IF of primary cultures revealed that high NuMA levels at mitotic spindle poles were significantly associated with a decreased proportion of cells in cytokinesis (p = 0.05), increased binucleation (p = 0.021) and multinucleation (p = 0.007), and aneuploidy (p = 0.008).NuMA is highly expressed in EOC tumours and high NuMA levels correlate with increases in mitotic defects and aneuploidy in primary cultures.

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NuMA expression in different ovarian tumour subtypes.Low and high NuMA staining patterns observed in cores from serous, mucinous, endometrial and clear cell carcinomas in a large scale ovarian TMA. Magnification x10.
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pone-0038945-g002: NuMA expression in different ovarian tumour subtypes.Low and high NuMA staining patterns observed in cores from serous, mucinous, endometrial and clear cell carcinomas in a large scale ovarian TMA. Magnification x10.

Mentions: Immunostaining indicated that NuMA expression in whole sections of normal ovarian tissue was variable, ranging from very low to weak levels in stromal and epithelial cell nuclei (n = 7) (Figure 1D). For the 5 cases of normal tissue with associated tumour tissue, a clear increase in NuMA expression in 40% (2/5) of tumour samples was identified. Up to 95% of nuclei in primary tumours displayed medium to strong NuMA staining (Figure S1a–d). NuMA was also observed at the spindle poles of mitotic tumour cells (Figure S1b, arrow). Analysis of a small in-house generated ovarian cancer TMA revealed that 95% of tumour cell nuclei were positively stained for NuMA in 24 of 25 cores. Strikingly, analysis of the data after application of a scoring system that subdivided samples into low NuMA (all negative, very weak and weak scores) and high NuMA levels (medium and high scores) showed that high NuMA levels were only detected in grade 3 tumours (33%; Figure 1E). Staining of a larger ovarian TMA confirmed low and high NuMA expression among the cores (Figure 2). Initial analysis of the overall data regardless of cancer subtype showed that high NuMA levels increased with tumour grade (from 20.3% for grade 1 tumours to 24.1% for grade 3 tumours, p = 0.0016) and disease stage (an increase from 19.6% for stage 1 to 33.3% for stage 4, p = 0.0077) (data not shown). Analysis of the continuous data for tumour grade and disease stage, which included subdivision into the cancer types for which enough data was available (serous, mucinous and endometrioid), indicated that NuMA expression correlated with increasing grade in the mucinous subtype (p = 0.009) (Figure 3A). A significant correlation with lymph node involvement (p = 0.03) (Figure 3B) and age (p = 0.04) (Figure 3C) was seen in all subtypes. Further analysis of the results as discontinuous data (low NuMA vs high NuMA) revealed additional statistically significant associations. The proportion of samples with high NuMA levels decreased with grade (p = 0.02) (Figure 3D) and increased in late disease stage in serous EOC (p = 0.04) (Figure 3E). In mucinous EOCs, NuMA levels increased with grade (p = 0.005) (Figure 3F). In late stage endometrioid EOCs, NuMA expression decreased (Figure 3G) and high NuMA decreased with an increase in tumour invasion status (p = 0.03) (all subtypes) (Figure 3H). Low NuMA expression was observed in the entire sample population of 13 clear cell EOCs regardless of disease stage.


NuMA overexpression in epithelial ovarian cancer.

Brüning-Richardson A, Bond J, Alsiary R, Richardson J, Cairns DA, McCormac L, Hutson R, Burns PA, Wilkinson N, Hall GD, Morrison EE, Bell SM - PLoS ONE (2012)

NuMA expression in different ovarian tumour subtypes.Low and high NuMA staining patterns observed in cores from serous, mucinous, endometrial and clear cell carcinomas in a large scale ovarian TMA. Magnification x10.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3375276&req=5

pone-0038945-g002: NuMA expression in different ovarian tumour subtypes.Low and high NuMA staining patterns observed in cores from serous, mucinous, endometrial and clear cell carcinomas in a large scale ovarian TMA. Magnification x10.
Mentions: Immunostaining indicated that NuMA expression in whole sections of normal ovarian tissue was variable, ranging from very low to weak levels in stromal and epithelial cell nuclei (n = 7) (Figure 1D). For the 5 cases of normal tissue with associated tumour tissue, a clear increase in NuMA expression in 40% (2/5) of tumour samples was identified. Up to 95% of nuclei in primary tumours displayed medium to strong NuMA staining (Figure S1a–d). NuMA was also observed at the spindle poles of mitotic tumour cells (Figure S1b, arrow). Analysis of a small in-house generated ovarian cancer TMA revealed that 95% of tumour cell nuclei were positively stained for NuMA in 24 of 25 cores. Strikingly, analysis of the data after application of a scoring system that subdivided samples into low NuMA (all negative, very weak and weak scores) and high NuMA levels (medium and high scores) showed that high NuMA levels were only detected in grade 3 tumours (33%; Figure 1E). Staining of a larger ovarian TMA confirmed low and high NuMA expression among the cores (Figure 2). Initial analysis of the overall data regardless of cancer subtype showed that high NuMA levels increased with tumour grade (from 20.3% for grade 1 tumours to 24.1% for grade 3 tumours, p = 0.0016) and disease stage (an increase from 19.6% for stage 1 to 33.3% for stage 4, p = 0.0077) (data not shown). Analysis of the continuous data for tumour grade and disease stage, which included subdivision into the cancer types for which enough data was available (serous, mucinous and endometrioid), indicated that NuMA expression correlated with increasing grade in the mucinous subtype (p = 0.009) (Figure 3A). A significant correlation with lymph node involvement (p = 0.03) (Figure 3B) and age (p = 0.04) (Figure 3C) was seen in all subtypes. Further analysis of the results as discontinuous data (low NuMA vs high NuMA) revealed additional statistically significant associations. The proportion of samples with high NuMA levels decreased with grade (p = 0.02) (Figure 3D) and increased in late disease stage in serous EOC (p = 0.04) (Figure 3E). In mucinous EOCs, NuMA levels increased with grade (p = 0.005) (Figure 3F). In late stage endometrioid EOCs, NuMA expression decreased (Figure 3G) and high NuMA decreased with an increase in tumour invasion status (p = 0.03) (all subtypes) (Figure 3H). Low NuMA expression was observed in the entire sample population of 13 clear cell EOCs regardless of disease stage.

Bottom Line: IHC revealed low to weak NuMA expression in normal tissues.NuMA expression decreased in late disease stage 4 endometrioid EOCs.IF of primary cultures revealed that high NuMA levels at mitotic spindle poles were significantly associated with a decreased proportion of cells in cytokinesis (p = 0.05), increased binucleation (p = 0.021) and multinucleation (p = 0.007), and aneuploidy (p = 0.008).NuMA is highly expressed in EOC tumours and high NuMA levels correlate with increases in mitotic defects and aneuploidy in primary cultures.

View Article: PubMed Central - PubMed

Affiliation: Section of Ophthalmology and Neuroscience, Leeds Institute of Molecular Medicine, St James's University Hospital, Leeds, United Kingdom. bgyar@leeds.ac.uk

ABSTRACT
Highly aneuploid tumours are common in epithelial ovarian cancers (EOC). We investigated whether NuMA expression was associated with this phenomenon.NuMA protein levels in normal and tumour tissues, ovarian cell lines and primary cultures of malignant cells derived from ovarian ascitic fluids were analysed by Affymetrix microarray analysis, immunoblotting, immunohistochemistry (IHC) and immunofluorescence (IF), with results correlated to associated clinical data. Aneuploidy status in primary cultures was determined by FACS analysis.Affymetrix microarray data indicated that NuMA was overexpressed in tumour tissue, primary cultures and cell lines compared to normal ovarian tissue. IHC revealed low to weak NuMA expression in normal tissues. Expression was upregulated in tumours, with a significant association with disease stage in mucinous EOC subtypes (p = 0.009), lymph node involvement (p = 0.03) and patient age (p = 0.04). Additional discontinuous data analysis revealed that high NuMA levels in tumours decreased with grade (p = 0.02) but increased with disease stage (p = 0.04) in serous EOC. NuMA expression decreased in late disease stage 4 endometrioid EOCs. High NuMA levels decreased with increased tumour invasion in all subtypes (p = 0.03). IF of primary cultures revealed that high NuMA levels at mitotic spindle poles were significantly associated with a decreased proportion of cells in cytokinesis (p = 0.05), increased binucleation (p = 0.021) and multinucleation (p = 0.007), and aneuploidy (p = 0.008).NuMA is highly expressed in EOC tumours and high NuMA levels correlate with increases in mitotic defects and aneuploidy in primary cultures.

Show MeSH
Related in: MedlinePlus