Limits...
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.

Show MeSH

Related in: MedlinePlus

NuMA expression in primary cultures is associated with mitotic stage, mitotic defects and aneuploidy.Strong nuclear NuMA expression was associated with (A) the lowest proportion of telophase cells and (B) the highest proportion of cells with a diffuse metaphase plate. Strong NuMA spindle pole labelling was associated with high rates of (C) binucleation, (D) multinucleation, (E) a diffuse metaphase plate and (F) the lowest proportion of cells in cytokinesis. (G) Intermediate spindle pole staining was associated with the lowest proportion of cells exhibiting a cytokinesis defect. (H) FACS analysis revealed that only cells with high levels of NuMA were aneuploid.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3375276&req=5

pone-0038945-g006: NuMA expression in primary cultures is associated with mitotic stage, mitotic defects and aneuploidy.Strong nuclear NuMA expression was associated with (A) the lowest proportion of telophase cells and (B) the highest proportion of cells with a diffuse metaphase plate. Strong NuMA spindle pole labelling was associated with high rates of (C) binucleation, (D) multinucleation, (E) a diffuse metaphase plate and (F) the lowest proportion of cells in cytokinesis. (G) Intermediate spindle pole staining was associated with the lowest proportion of cells exhibiting a cytokinesis defect. (H) FACS analysis revealed that only cells with high levels of NuMA were aneuploid.

Mentions: A comparison of NuMA staining profiles to mitotic data was performed. High levels of NuMA in interphase nuclei were significantly associated with a small proportion of cells in telophase (p = 0.012) and a high proportion of cells displaying a loose metaphase plate (p = 0.032) (Figure 6A and B). High levels of NuMA at mitotic spindle poles significantly correlated with high rates of binucleation (p = 0.021) and multinucleation (p = 0.007) (Figure 6C and D) and again with loose metaphase plates (p = 0.041) (Figure 6E). Low levels of NuMA at spindle poles significantly correlated with a high proportion of cells in cytokinesis (p = 0.05) (Figure 6F). Interestingly, intermediate levels of NuMA at the spindle poles significantly correlated with low error rates in cytokinesis. (p = 0.01) (Figure 6G). To further verify the data observed in our immunofluorescence analysis of the primary cultures we immunostained whole sections of solid tumour tissue from the 23 patients whose ascitic fluids were used to generate the cultures. This confirmed that solid tumours displaying high levels of NuMA staining generated primary ascitic cultures where cells displayed high NuMA expression levels (Figure S1 a–d). To ascertain that the observed interphase and mitotic defects were also a common feature of the associated tumour tissues we analysed 23 tumour sections available to us by NuMA immunostaining and H&E IHC staining. We noted that the same interphase and mitotic defects were present in these tumours as in the ascites samples. Representative images and a summary of the defects observed among the samples are shown in Figure S2A–D. Furthermore, cell cultures with a high mitotic index were found to display high mitotic activity in the associated tumour tissue (Figure S3). Finally, we explored the relationship between NuMA expression levels in primary cultures and patient survival. This suggested that patients with moderate to strong NuMA immunostaining at the spindle poles in primary cultures have a lower survival rate than patients with weak NuMA immunostaining, though this preliminary data did not reach statistical significance (Figure S4), perhaps due to low sample size.


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 primary cultures is associated with mitotic stage, mitotic defects and aneuploidy.Strong nuclear NuMA expression was associated with (A) the lowest proportion of telophase cells and (B) the highest proportion of cells with a diffuse metaphase plate. Strong NuMA spindle pole labelling was associated with high rates of (C) binucleation, (D) multinucleation, (E) a diffuse metaphase plate and (F) the lowest proportion of cells in cytokinesis. (G) Intermediate spindle pole staining was associated with the lowest proportion of cells exhibiting a cytokinesis defect. (H) FACS analysis revealed that only cells with high levels of NuMA were aneuploid.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038945-g006: NuMA expression in primary cultures is associated with mitotic stage, mitotic defects and aneuploidy.Strong nuclear NuMA expression was associated with (A) the lowest proportion of telophase cells and (B) the highest proportion of cells with a diffuse metaphase plate. Strong NuMA spindle pole labelling was associated with high rates of (C) binucleation, (D) multinucleation, (E) a diffuse metaphase plate and (F) the lowest proportion of cells in cytokinesis. (G) Intermediate spindle pole staining was associated with the lowest proportion of cells exhibiting a cytokinesis defect. (H) FACS analysis revealed that only cells with high levels of NuMA were aneuploid.
Mentions: A comparison of NuMA staining profiles to mitotic data was performed. High levels of NuMA in interphase nuclei were significantly associated with a small proportion of cells in telophase (p = 0.012) and a high proportion of cells displaying a loose metaphase plate (p = 0.032) (Figure 6A and B). High levels of NuMA at mitotic spindle poles significantly correlated with high rates of binucleation (p = 0.021) and multinucleation (p = 0.007) (Figure 6C and D) and again with loose metaphase plates (p = 0.041) (Figure 6E). Low levels of NuMA at spindle poles significantly correlated with a high proportion of cells in cytokinesis (p = 0.05) (Figure 6F). Interestingly, intermediate levels of NuMA at the spindle poles significantly correlated with low error rates in cytokinesis. (p = 0.01) (Figure 6G). To further verify the data observed in our immunofluorescence analysis of the primary cultures we immunostained whole sections of solid tumour tissue from the 23 patients whose ascitic fluids were used to generate the cultures. This confirmed that solid tumours displaying high levels of NuMA staining generated primary ascitic cultures where cells displayed high NuMA expression levels (Figure S1 a–d). To ascertain that the observed interphase and mitotic defects were also a common feature of the associated tumour tissues we analysed 23 tumour sections available to us by NuMA immunostaining and H&E IHC staining. We noted that the same interphase and mitotic defects were present in these tumours as in the ascites samples. Representative images and a summary of the defects observed among the samples are shown in Figure S2A–D. Furthermore, cell cultures with a high mitotic index were found to display high mitotic activity in the associated tumour tissue (Figure S3). Finally, we explored the relationship between NuMA expression levels in primary cultures and patient survival. This suggested that patients with moderate to strong NuMA immunostaining at the spindle poles in primary cultures have a lower survival rate than patients with weak NuMA immunostaining, though this preliminary data did not reach statistical significance (Figure S4), perhaps due to low sample size.

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