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WWOX protein expression varies among ovarian carcinoma histotypes and correlates with less favorable outcome.

Nunez MI, Rosen DG, Ludes-Meyers JH, Abba MC, Kil H, Page R, Klein-Szanto AJ, Godwin AK, Liu J, Mills GB, Aldaz CM - BMC Cancer (2005)

Bottom Line: Immunoblotting analysis from normal ovarian samples demonstrated consistently strong WWOX expression while 37% ovarian carcinomas showed reduced or undetectable WWOX protein expression levels.The two histotypes showing significant loss of WWOX expression were of the Mucinous (70%) and Clear Cell (42%) types.It was also observed that subsets of ovarian tumors demonstrated loss of WWOX expression and is potentially associated with patient outcome.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Carcinogenesis, The University of Texas M,D, Anderson Cancer Center, Science Park-Research Division, Smithville TX, USA. minunez@mdanderson.org <minunez@mdanderson.org>

ABSTRACT

Background: The putative tumor suppressor WWOX gene spans the common chromosomal fragile site 16D (FRA16D) at chromosome area 16q23.3-24.1. This region is a frequent target for loss of heterozygosity and chromosomal rearrangement in ovarian, breast, hepatocellular, prostate carcinomas and other neoplasias. The goal of these studies was to evaluate WWOX protein expression levels in ovarian carcinomas to determine if they correlated with clinico-pathological parameters, thus providing additional support for WWOX functioning as a tumor suppressor.

Methods: We performed WWOX protein expression analyses by means of immunobloting and immunohistochemistry on normal ovaries and specific human ovarian carcinoma Tissue Microarrays (n = 444). Univariate analysis of clinical-pathological parameters based on WWOX staining was determined by chi2 test with Yates' correction. The basic significance level was fixed at p < 0.05.

Results: Immunoblotting analysis from normal ovarian samples demonstrated consistently strong WWOX expression while 37% ovarian carcinomas showed reduced or undetectable WWOX protein expression levels. The immunohistochemistry of normal human ovarian tissue sections confirmed strong WWOX expression in ovarian surface epithelial cells and in epithelial inclusion cysts within the cortex. Out of 444 ovarian carcinoma samples analyzed 30% of tumors showed lack of or barely detectable WWOX expression. The remaining ovarian carcinomas (70%) stained moderately to strongly positive for this protein. The two histotypes showing significant loss of WWOX expression were of the Mucinous (70%) and Clear Cell (42%) types. Reduced WWOX expression demonstrated a significant association with clinical Stage IV (FIGO) (p = 0.007), negative Progesterone Receptor (PR) status (p = 0.008) and shorter overall survival (p = 0.03).

Conclusion: These data indicate that WWOX protein expression is highly variable among ovarian carcinoma histotypes. It was also observed that subsets of ovarian tumors demonstrated loss of WWOX expression and is potentially associated with patient outcome.

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WWOX protein expression in normal ovary and ovarian adenocarcinomas as determined by immunoblot analysis. A) Total protein extracts from PEO1 cell lines transfected with an empty vector or a WWOX expressing vector were analyzed by immunobloting with the anti-WWOX antibody. Note: no immunoreactive bands are observed in the vector transfected cell line. B) WWOX protein expression was determined by immunoblotting of total protein extracts from 38 ovarian carcinoma samples. Five normal ovarian tissue extracts are shown on the first five lanes. Quantitation of WWOX protein expression. Autoradiographs of WWOX and actin were digitized using the Kodak digital science Image Station 440CF. WWOX expression in each sample was normalizated to actin to correct for loading differences. In turn these numbers obtained from each tumor were normalized and expressed as relative to the normal ovarian values (i.e. Relative Expression).
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Figure 1: WWOX protein expression in normal ovary and ovarian adenocarcinomas as determined by immunoblot analysis. A) Total protein extracts from PEO1 cell lines transfected with an empty vector or a WWOX expressing vector were analyzed by immunobloting with the anti-WWOX antibody. Note: no immunoreactive bands are observed in the vector transfected cell line. B) WWOX protein expression was determined by immunoblotting of total protein extracts from 38 ovarian carcinoma samples. Five normal ovarian tissue extracts are shown on the first five lanes. Quantitation of WWOX protein expression. Autoradiographs of WWOX and actin were digitized using the Kodak digital science Image Station 440CF. WWOX expression in each sample was normalizated to actin to correct for loading differences. In turn these numbers obtained from each tumor were normalized and expressed as relative to the normal ovarian values (i.e. Relative Expression).

Mentions: Immunoblotting analysis demonstrated that full length WWOX protein was the predominant WWOX isoform expressed in both normal ovary and ovarian carcinomas (Figure 1B). The anti-WWOX antibody used was raised against the WW domains [2] and should detect all potential WWOX isoforms. The specificity of this WWOX antibody in the immunoblotting analyses was demonstrated by observing no immunoreactive products in cell extracts from the PEO1 ovarian carcinoma cell line used as negative control (Figure 1A). In addition, as a positive control PEO1 cells were stably transfected with a WWOX expression vector (Figure 1A).


WWOX protein expression varies among ovarian carcinoma histotypes and correlates with less favorable outcome.

Nunez MI, Rosen DG, Ludes-Meyers JH, Abba MC, Kil H, Page R, Klein-Szanto AJ, Godwin AK, Liu J, Mills GB, Aldaz CM - BMC Cancer (2005)

WWOX protein expression in normal ovary and ovarian adenocarcinomas as determined by immunoblot analysis. A) Total protein extracts from PEO1 cell lines transfected with an empty vector or a WWOX expressing vector were analyzed by immunobloting with the anti-WWOX antibody. Note: no immunoreactive bands are observed in the vector transfected cell line. B) WWOX protein expression was determined by immunoblotting of total protein extracts from 38 ovarian carcinoma samples. Five normal ovarian tissue extracts are shown on the first five lanes. Quantitation of WWOX protein expression. Autoradiographs of WWOX and actin were digitized using the Kodak digital science Image Station 440CF. WWOX expression in each sample was normalizated to actin to correct for loading differences. In turn these numbers obtained from each tumor were normalized and expressed as relative to the normal ovarian values (i.e. Relative Expression).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1173095&req=5

Figure 1: WWOX protein expression in normal ovary and ovarian adenocarcinomas as determined by immunoblot analysis. A) Total protein extracts from PEO1 cell lines transfected with an empty vector or a WWOX expressing vector were analyzed by immunobloting with the anti-WWOX antibody. Note: no immunoreactive bands are observed in the vector transfected cell line. B) WWOX protein expression was determined by immunoblotting of total protein extracts from 38 ovarian carcinoma samples. Five normal ovarian tissue extracts are shown on the first five lanes. Quantitation of WWOX protein expression. Autoradiographs of WWOX and actin were digitized using the Kodak digital science Image Station 440CF. WWOX expression in each sample was normalizated to actin to correct for loading differences. In turn these numbers obtained from each tumor were normalized and expressed as relative to the normal ovarian values (i.e. Relative Expression).
Mentions: Immunoblotting analysis demonstrated that full length WWOX protein was the predominant WWOX isoform expressed in both normal ovary and ovarian carcinomas (Figure 1B). The anti-WWOX antibody used was raised against the WW domains [2] and should detect all potential WWOX isoforms. The specificity of this WWOX antibody in the immunoblotting analyses was demonstrated by observing no immunoreactive products in cell extracts from the PEO1 ovarian carcinoma cell line used as negative control (Figure 1A). In addition, as a positive control PEO1 cells were stably transfected with a WWOX expression vector (Figure 1A).

Bottom Line: Immunoblotting analysis from normal ovarian samples demonstrated consistently strong WWOX expression while 37% ovarian carcinomas showed reduced or undetectable WWOX protein expression levels.The two histotypes showing significant loss of WWOX expression were of the Mucinous (70%) and Clear Cell (42%) types.It was also observed that subsets of ovarian tumors demonstrated loss of WWOX expression and is potentially associated with patient outcome.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Carcinogenesis, The University of Texas M,D, Anderson Cancer Center, Science Park-Research Division, Smithville TX, USA. minunez@mdanderson.org <minunez@mdanderson.org>

ABSTRACT

Background: The putative tumor suppressor WWOX gene spans the common chromosomal fragile site 16D (FRA16D) at chromosome area 16q23.3-24.1. This region is a frequent target for loss of heterozygosity and chromosomal rearrangement in ovarian, breast, hepatocellular, prostate carcinomas and other neoplasias. The goal of these studies was to evaluate WWOX protein expression levels in ovarian carcinomas to determine if they correlated with clinico-pathological parameters, thus providing additional support for WWOX functioning as a tumor suppressor.

Methods: We performed WWOX protein expression analyses by means of immunobloting and immunohistochemistry on normal ovaries and specific human ovarian carcinoma Tissue Microarrays (n = 444). Univariate analysis of clinical-pathological parameters based on WWOX staining was determined by chi2 test with Yates' correction. The basic significance level was fixed at p < 0.05.

Results: Immunoblotting analysis from normal ovarian samples demonstrated consistently strong WWOX expression while 37% ovarian carcinomas showed reduced or undetectable WWOX protein expression levels. The immunohistochemistry of normal human ovarian tissue sections confirmed strong WWOX expression in ovarian surface epithelial cells and in epithelial inclusion cysts within the cortex. Out of 444 ovarian carcinoma samples analyzed 30% of tumors showed lack of or barely detectable WWOX expression. The remaining ovarian carcinomas (70%) stained moderately to strongly positive for this protein. The two histotypes showing significant loss of WWOX expression were of the Mucinous (70%) and Clear Cell (42%) types. Reduced WWOX expression demonstrated a significant association with clinical Stage IV (FIGO) (p = 0.007), negative Progesterone Receptor (PR) status (p = 0.008) and shorter overall survival (p = 0.03).

Conclusion: These data indicate that WWOX protein expression is highly variable among ovarian carcinoma histotypes. It was also observed that subsets of ovarian tumors demonstrated loss of WWOX expression and is potentially associated with patient outcome.

Show MeSH
Related in: MedlinePlus