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NUCKS overexpression in breast cancer.

Drosos Y, Kouloukoussa M, Østvold AC, Grundt K, Goutas N, Vlachodimitropoulos D, Havaki S, Kollia P, Kittas C, Marinos E, Aleporou-Marinou V - Cancer Cell Int. (2009)

Bottom Line: This increase was also confirmed by Western immunoblot analysis.Although NUCKS is a cell cycle related protein, its expression does not correlate with Ki67 expression, neither in tissue sections nor in primary cell cultures.In particular, the NUCKS overexpression in ADH and DCIS indicates a significant role of this protein in neoplastic progression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Genetics and Biotechnology, Faculty of Biology, University of Athens, Panepistimioupoli, 15701 Ilissia, Greece. ydrosos@biol.uoa.gr

ABSTRACT

Background: NUCKS (Nuclear, Casein Kinase and Cyclin-dependent Kinase Substrate) is a nuclear, DNA-binding and highly phosphorylated protein. A number of reports show that NUCKS is highly expressed on the level of mRNA in several human cancers, including breast cancer. In this work, NUCKS expression on both RNA and protein levels was studied in breast tissue biopsies consisted of invasive carcinomas, intraductal proliferative lesions, benign epithelial proliferations and fibroadenomas, as well as in primary cultures derived from the above biopsies. Specifically, in order to evaluate the level of NUCKS protein in correlation with the histopathological features of breast disease, immunohistochemistry was employed on paraffin sections of breast biopsies of the above types. In addition, NUCKS expression was studied by means of Reverse Transcription PCR (RT-PCR), real-time PCR (qRT-PCR) and Western immunoblot analyses in the primary cell cultures developed from the same biopsies.

Results: The immunohistochemical Results showed intense NUCKS staining mostly in grade I and II breast carcinomas compared to normal tissues. Furthermore, NUCKS was moderate expressed in benign epithelial proliferations, such as adenosis and sclerosing adenosis, and highly expressed in intraductal lesions, specifically in ductal carcinomas in situ (DCIS). It is worth noting that all the fibroadenoma tissues examined were negative for NUCKS staining. RT-PCR and qRT-PCR showed an increase of NUCKS expression in cells derived from primary cultures of proliferative lesions and cancerous tissues compared to the ones derived from normal breast tissues and fibroadenomas. This increase was also confirmed by Western immunoblot analysis. Although NUCKS is a cell cycle related protein, its expression does not correlate with Ki67 expression, neither in tissue sections nor in primary cell cultures.

Conclusion: The results show overexpression of the NUCKS protein in a number of non malignant breast lesions and cancerous tissues. In particular, the NUCKS overexpression in ADH and DCIS indicates a significant role of this protein in neoplastic progression.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemical staining of DCIS and IDC with anti-NUCKS and anti-ki67 antibody. Tissue sections stained with anti-NUCKS (A-C) and corresponding sections stained with anti-ki67 (D-F) antibody. (A), (D), Strong immunoreaction for NUCKS and low for Ki67 in DCIS. (B), (E) Strong immunoreaction for NUCKS and low for Ki67 in IDC, grade II. (C), (F) IDC, grade III negative staining for NUCKS and strong staining for Ki67. Scale bar, 50 μm.
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Figure 2: Immunohistochemical staining of DCIS and IDC with anti-NUCKS and anti-ki67 antibody. Tissue sections stained with anti-NUCKS (A-C) and corresponding sections stained with anti-ki67 (D-F) antibody. (A), (D), Strong immunoreaction for NUCKS and low for Ki67 in DCIS. (B), (E) Strong immunoreaction for NUCKS and low for Ki67 in IDC, grade II. (C), (F) IDC, grade III negative staining for NUCKS and strong staining for Ki67. Scale bar, 50 μm.

Mentions: All the fibroadenoma cases (13/13) as well as the normal tissues that were examined, stained negatively for NUCKS (Table 1, Figures 1A, 1B). In the benign proliferative lesions examined, NUCKS was expressed in 17/23 (85%) cases of adenosis and 10/11 (91%) cases of sclerosing adenosis (Table 1, Figure 1C) exhibiting moderate staining. In the intraductal proliferative lesions, NUCKS was expressed in 13/16 (81%) cases of usual ductal hyperplasia (UDH), in 6/6 (100%) cases of atypical ductal hyperplasia (ADH) and in 15/16 (94%) cases of DCIS, exhibiting intense staining in the majority of ADH and DCIS (83% and 82% of the cases respectively stained with a score of +2/+3) (Table 1, Figures 1D, 2A).


NUCKS overexpression in breast cancer.

Drosos Y, Kouloukoussa M, Østvold AC, Grundt K, Goutas N, Vlachodimitropoulos D, Havaki S, Kollia P, Kittas C, Marinos E, Aleporou-Marinou V - Cancer Cell Int. (2009)

Immunohistochemical staining of DCIS and IDC with anti-NUCKS and anti-ki67 antibody. Tissue sections stained with anti-NUCKS (A-C) and corresponding sections stained with anti-ki67 (D-F) antibody. (A), (D), Strong immunoreaction for NUCKS and low for Ki67 in DCIS. (B), (E) Strong immunoreaction for NUCKS and low for Ki67 in IDC, grade II. (C), (F) IDC, grade III negative staining for NUCKS and strong staining for Ki67. Scale bar, 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2743642&req=5

Figure 2: Immunohistochemical staining of DCIS and IDC with anti-NUCKS and anti-ki67 antibody. Tissue sections stained with anti-NUCKS (A-C) and corresponding sections stained with anti-ki67 (D-F) antibody. (A), (D), Strong immunoreaction for NUCKS and low for Ki67 in DCIS. (B), (E) Strong immunoreaction for NUCKS and low for Ki67 in IDC, grade II. (C), (F) IDC, grade III negative staining for NUCKS and strong staining for Ki67. Scale bar, 50 μm.
Mentions: All the fibroadenoma cases (13/13) as well as the normal tissues that were examined, stained negatively for NUCKS (Table 1, Figures 1A, 1B). In the benign proliferative lesions examined, NUCKS was expressed in 17/23 (85%) cases of adenosis and 10/11 (91%) cases of sclerosing adenosis (Table 1, Figure 1C) exhibiting moderate staining. In the intraductal proliferative lesions, NUCKS was expressed in 13/16 (81%) cases of usual ductal hyperplasia (UDH), in 6/6 (100%) cases of atypical ductal hyperplasia (ADH) and in 15/16 (94%) cases of DCIS, exhibiting intense staining in the majority of ADH and DCIS (83% and 82% of the cases respectively stained with a score of +2/+3) (Table 1, Figures 1D, 2A).

Bottom Line: This increase was also confirmed by Western immunoblot analysis.Although NUCKS is a cell cycle related protein, its expression does not correlate with Ki67 expression, neither in tissue sections nor in primary cell cultures.In particular, the NUCKS overexpression in ADH and DCIS indicates a significant role of this protein in neoplastic progression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Genetics and Biotechnology, Faculty of Biology, University of Athens, Panepistimioupoli, 15701 Ilissia, Greece. ydrosos@biol.uoa.gr

ABSTRACT

Background: NUCKS (Nuclear, Casein Kinase and Cyclin-dependent Kinase Substrate) is a nuclear, DNA-binding and highly phosphorylated protein. A number of reports show that NUCKS is highly expressed on the level of mRNA in several human cancers, including breast cancer. In this work, NUCKS expression on both RNA and protein levels was studied in breast tissue biopsies consisted of invasive carcinomas, intraductal proliferative lesions, benign epithelial proliferations and fibroadenomas, as well as in primary cultures derived from the above biopsies. Specifically, in order to evaluate the level of NUCKS protein in correlation with the histopathological features of breast disease, immunohistochemistry was employed on paraffin sections of breast biopsies of the above types. In addition, NUCKS expression was studied by means of Reverse Transcription PCR (RT-PCR), real-time PCR (qRT-PCR) and Western immunoblot analyses in the primary cell cultures developed from the same biopsies.

Results: The immunohistochemical Results showed intense NUCKS staining mostly in grade I and II breast carcinomas compared to normal tissues. Furthermore, NUCKS was moderate expressed in benign epithelial proliferations, such as adenosis and sclerosing adenosis, and highly expressed in intraductal lesions, specifically in ductal carcinomas in situ (DCIS). It is worth noting that all the fibroadenoma tissues examined were negative for NUCKS staining. RT-PCR and qRT-PCR showed an increase of NUCKS expression in cells derived from primary cultures of proliferative lesions and cancerous tissues compared to the ones derived from normal breast tissues and fibroadenomas. This increase was also confirmed by Western immunoblot analysis. Although NUCKS is a cell cycle related protein, its expression does not correlate with Ki67 expression, neither in tissue sections nor in primary cell cultures.

Conclusion: The results show overexpression of the NUCKS protein in a number of non malignant breast lesions and cancerous tissues. In particular, the NUCKS overexpression in ADH and DCIS indicates a significant role of this protein in neoplastic progression.

No MeSH data available.


Related in: MedlinePlus