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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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Mitotic and interphase defects in primary cultures.A. Mitotic defects observed at metaphase included (a) asymmetrical bipolar spindles (b) multiple spindle poles, (c) tripolar spindles, (d) bipolar spindles with multiple spindle poles (e) cells with misplaced and misaligned spindles (cell periphery outlined). B. Mitotic defects observed at anaphase included lagging chromosomes and anaphase bridging (a -c). C. Mitotic defects observed during telophase and cytokinesis included (a) abnormal spindles, (b) abnormal cytokinesis with micronuclei formation, (c), chromatid bridging and (d) formation of unevenly sized daughter cells. D. Examples of (a) binucleation, (b) multinucleation and (c) micronuclei in primary cultures. Green – NuMA, red – α-tubulin, blue – DAPI in all panels. Scale bar in A and B = 5 µm. Scale bar in C and D = 10 µm. E. Pie chart illustrating the overall frequency of mitotic defects observed in the primary cultures.
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pone-0038945-g005: Mitotic and interphase defects in primary cultures.A. Mitotic defects observed at metaphase included (a) asymmetrical bipolar spindles (b) multiple spindle poles, (c) tripolar spindles, (d) bipolar spindles with multiple spindle poles (e) cells with misplaced and misaligned spindles (cell periphery outlined). B. Mitotic defects observed at anaphase included lagging chromosomes and anaphase bridging (a -c). C. Mitotic defects observed during telophase and cytokinesis included (a) abnormal spindles, (b) abnormal cytokinesis with micronuclei formation, (c), chromatid bridging and (d) formation of unevenly sized daughter cells. D. Examples of (a) binucleation, (b) multinucleation and (c) micronuclei in primary cultures. Green – NuMA, red – α-tubulin, blue – DAPI in all panels. Scale bar in A and B = 5 µm. Scale bar in C and D = 10 µm. E. Pie chart illustrating the overall frequency of mitotic defects observed in the primary cultures.

Mentions: Staining intensities differed amongst the samples (Figure 4). It was noted that some samples were composed of cells with low level NuMA staining in interphase nuclei and at mitotic spindle poles (Figure 4A, a–d and i–l; 4B, C) and some samples were composed of cells with high NuMA staining intensities (Figure 4A, e–h and m–p; Figure 4B and C). We also noted that each sample differed with regards to the proportion of cells at each mitotic stage and that they exhibited a wide range of mitotic defects. These included metaphase defects such as tripolar and multipolar spindles (Figure 5A, a–d) and spindle misalignment (Figure 5A, e); anaphase defects such as lagging chromosomes and chromosomal bridging (Figure 5B, a–c); and cytokinetic defects including chromatid bridging and the generation of unevenly sized daughter cells (Figure 5C, a–d). Binucleation, multinucleation and the presence of micronuclei were common defects in interphase cells (Figure 5D, a–c). The examples shown in Figure 5E were the most common mitotic errors observed.


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)

Mitotic and interphase defects in primary cultures.A. Mitotic defects observed at metaphase included (a) asymmetrical bipolar spindles (b) multiple spindle poles, (c) tripolar spindles, (d) bipolar spindles with multiple spindle poles (e) cells with misplaced and misaligned spindles (cell periphery outlined). B. Mitotic defects observed at anaphase included lagging chromosomes and anaphase bridging (a -c). C. Mitotic defects observed during telophase and cytokinesis included (a) abnormal spindles, (b) abnormal cytokinesis with micronuclei formation, (c), chromatid bridging and (d) formation of unevenly sized daughter cells. D. Examples of (a) binucleation, (b) multinucleation and (c) micronuclei in primary cultures. Green – NuMA, red – α-tubulin, blue – DAPI in all panels. Scale bar in A and B = 5 µm. Scale bar in C and D = 10 µm. E. Pie chart illustrating the overall frequency of mitotic defects observed in the primary cultures.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038945-g005: Mitotic and interphase defects in primary cultures.A. Mitotic defects observed at metaphase included (a) asymmetrical bipolar spindles (b) multiple spindle poles, (c) tripolar spindles, (d) bipolar spindles with multiple spindle poles (e) cells with misplaced and misaligned spindles (cell periphery outlined). B. Mitotic defects observed at anaphase included lagging chromosomes and anaphase bridging (a -c). C. Mitotic defects observed during telophase and cytokinesis included (a) abnormal spindles, (b) abnormal cytokinesis with micronuclei formation, (c), chromatid bridging and (d) formation of unevenly sized daughter cells. D. Examples of (a) binucleation, (b) multinucleation and (c) micronuclei in primary cultures. Green – NuMA, red – α-tubulin, blue – DAPI in all panels. Scale bar in A and B = 5 µm. Scale bar in C and D = 10 µm. E. Pie chart illustrating the overall frequency of mitotic defects observed in the primary cultures.
Mentions: Staining intensities differed amongst the samples (Figure 4). It was noted that some samples were composed of cells with low level NuMA staining in interphase nuclei and at mitotic spindle poles (Figure 4A, a–d and i–l; 4B, C) and some samples were composed of cells with high NuMA staining intensities (Figure 4A, e–h and m–p; Figure 4B and C). We also noted that each sample differed with regards to the proportion of cells at each mitotic stage and that they exhibited a wide range of mitotic defects. These included metaphase defects such as tripolar and multipolar spindles (Figure 5A, a–d) and spindle misalignment (Figure 5A, e); anaphase defects such as lagging chromosomes and chromosomal bridging (Figure 5B, a–c); and cytokinetic defects including chromatid bridging and the generation of unevenly sized daughter cells (Figure 5C, a–d). Binucleation, multinucleation and the presence of micronuclei were common defects in interphase cells (Figure 5D, a–c). The examples shown in Figure 5E were the most common mitotic errors observed.

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