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Localization of hRad9 in breast cancer.

Chan V, Khoo US, Wong MS, Lau K, Suen D, Li G, Kwong A, Chan TK - BMC Cancer (2008)

Bottom Line: We have previously shown that the mRNA up-regulation correlated with tumor size and local recurrence.Localisation of hRad9 protein were performed on paired tumor and normal breast tissues.This nuclear protein existed in hyperphosphorylated forms which may be those of the hRad9-hRad1-hHus1 complex.

View Article: PubMed Central - HTML - PubMed

Affiliation: University Department of Medicine, Queen Mary Hospital, Hong Kong, China. vnychana@hkucc.hku.hk

ABSTRACT

Background: hRad9 is a cell cycle checkpoint gene that is up-regulated in breast cancer. We have previously shown that the mRNA up-regulation correlated with tumor size and local recurrence. Immunohistochemical studies were made to better define the role of hRad9 in breast carcinogenesis.

Methods: Localisation of hRad9 protein were performed on paired tumor and normal breast tissues. Immunoblotting with and without dephosphorylation was used to define the protein isolated from breast cancer cells.

Results: Increased hRad9 protein was observed in breast cancer cells nucleus compared to non-tumor epithelium. This nuclear protein existed in hyperphosphorylated forms which may be those of the hRad9-hRad1-hHus1 complex.

Conclusion: Finding of hyperphosphorylated forms of hRad9 in the nucleus of cancer cells is in keeping with its function in ameliorating DNA instability, whereby it inadvertently assists tumor growth.

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Related in: MedlinePlus

Western blot showing hRad9 protein extracted from MDA231 breast cancer cells, breast tumor sample with nuclear staining of hRad9 (A) and a control tumor sample with no nuclear staining (B). The relative position for molecular weights (in KDa) are indicated on the right. Protein extracts were treated with λ phosphatase (100 and 500 units respectively).
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Figure 2: Western blot showing hRad9 protein extracted from MDA231 breast cancer cells, breast tumor sample with nuclear staining of hRad9 (A) and a control tumor sample with no nuclear staining (B). The relative position for molecular weights (in KDa) are indicated on the right. Protein extracts were treated with λ phosphatase (100 and 500 units respectively).

Mentions: Western blot analysis showed at least four different species of hRad9 protein in a breast tumor sample with positive nuclear staining, with molecular weight ranging from 65 to 45 KDa. Whereas in MDA-231 breast cancer cell line, the hRad9 protein was mainly 50 and 45 KDa. Dephosphorylation of hRad9 protein extracted from both the tumor sample with nuclear staining and the MDA-231 cancer cell line with λ phosphatase yielded an additional 48 KDa species. The control tumor sample (which had no nuclear hRad9 staining), did not produce any 48 KDa species even upon treatment with 500 units of λ phosphatase (Figure 2).


Localization of hRad9 in breast cancer.

Chan V, Khoo US, Wong MS, Lau K, Suen D, Li G, Kwong A, Chan TK - BMC Cancer (2008)

Western blot showing hRad9 protein extracted from MDA231 breast cancer cells, breast tumor sample with nuclear staining of hRad9 (A) and a control tumor sample with no nuclear staining (B). The relative position for molecular weights (in KDa) are indicated on the right. Protein extracts were treated with λ phosphatase (100 and 500 units respectively).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2483722&req=5

Figure 2: Western blot showing hRad9 protein extracted from MDA231 breast cancer cells, breast tumor sample with nuclear staining of hRad9 (A) and a control tumor sample with no nuclear staining (B). The relative position for molecular weights (in KDa) are indicated on the right. Protein extracts were treated with λ phosphatase (100 and 500 units respectively).
Mentions: Western blot analysis showed at least four different species of hRad9 protein in a breast tumor sample with positive nuclear staining, with molecular weight ranging from 65 to 45 KDa. Whereas in MDA-231 breast cancer cell line, the hRad9 protein was mainly 50 and 45 KDa. Dephosphorylation of hRad9 protein extracted from both the tumor sample with nuclear staining and the MDA-231 cancer cell line with λ phosphatase yielded an additional 48 KDa species. The control tumor sample (which had no nuclear hRad9 staining), did not produce any 48 KDa species even upon treatment with 500 units of λ phosphatase (Figure 2).

Bottom Line: We have previously shown that the mRNA up-regulation correlated with tumor size and local recurrence.Localisation of hRad9 protein were performed on paired tumor and normal breast tissues.This nuclear protein existed in hyperphosphorylated forms which may be those of the hRad9-hRad1-hHus1 complex.

View Article: PubMed Central - HTML - PubMed

Affiliation: University Department of Medicine, Queen Mary Hospital, Hong Kong, China. vnychana@hkucc.hku.hk

ABSTRACT

Background: hRad9 is a cell cycle checkpoint gene that is up-regulated in breast cancer. We have previously shown that the mRNA up-regulation correlated with tumor size and local recurrence. Immunohistochemical studies were made to better define the role of hRad9 in breast carcinogenesis.

Methods: Localisation of hRad9 protein were performed on paired tumor and normal breast tissues. Immunoblotting with and without dephosphorylation was used to define the protein isolated from breast cancer cells.

Results: Increased hRad9 protein was observed in breast cancer cells nucleus compared to non-tumor epithelium. This nuclear protein existed in hyperphosphorylated forms which may be those of the hRad9-hRad1-hHus1 complex.

Conclusion: Finding of hyperphosphorylated forms of hRad9 in the nucleus of cancer cells is in keeping with its function in ameliorating DNA instability, whereby it inadvertently assists tumor growth.

Show MeSH
Related in: MedlinePlus