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Histone H1 phosphorylation in breast cancer.

Harshman SW, Hoover ME, Huang C, Branson OE, Chaney SB, Cheney CM, Rosol TJ, Shapiro CL, Wysocki VH, Huebner K, Freitas MA - J. Proteome Res. (2014)

Bottom Line: The results show that the extent of H1 phosphorylation can distinguish between the different cell lines.The results suggest that the phosphorylation at threonine 146 is found on both histone H1.2 and histone H1.4.The data show that histone H1 phosphorylation can increase and decrease in response to extracellular stimuli.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Virology, Immunology and Medical Genetics, ‡Comprehensive Cancer Center, §Department of Chemistry & Biochemistry, ⊥Veterinary Biosciences, College of Veterinary Medicine, and ∥Department of Internal Medicine, Division of Hematology, The Ohio State University , Columbus, Ohio 43210, United States.

ABSTRACT
Breast cancer is the second leading cause of cancer-related deaths in women. The need for new clinical biomarkers in breast cancer is necessary to further predict prognosis and therapeutic response. In this article, the LC-MS histone H1 phosphorylation profiles were established for three distinct breast cancer cell lines. The results show that the extent of H1 phosphorylation can distinguish between the different cell lines. The histone H1 from the metastatic cell line, MDA-MB-231, was subjected to chemical derivitization and LC-MS/MS analysis. The results suggest that the phosphorylation at threonine 146 is found on both histone H1.2 and histone H1.4. Cell lines were then treated with an extracellular stimulus, estradiol or kinase inhibitor LY294002, to monitor changes in histone H1 phosphorylation. The data show that histone H1 phosphorylation can increase and decrease in response to extracellular stimuli. Finally, primary breast tissues were stained for the histone H1 phosphorylation at threonine 146. Variable staining patterns across tumor grades and subtypes were observed with pT146 labeling correlating with tumor grade. These results establish the potential for histone H1 phosphorylation at threonine 146 as a clinical biomarker in breast cancer.

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Related in: MedlinePlus

Immunohistochemicalstaining of primary breast tumorsfor Hematoxylin, eosin (H&E) and pT146: (A) grade I Luminal A,(B) grade II Luminal A, (C) grade III Luminal A, (D) grade II LuminalB, (E) grade III Luminal B, (F) grade II Her2, (G) grade III Her2,(H) grade II triple negative, (I) grade III triple negative. Datashow specific staining patterns based upon grade and tissue subtype.
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fig6: Immunohistochemicalstaining of primary breast tumorsfor Hematoxylin, eosin (H&E) and pT146: (A) grade I Luminal A,(B) grade II Luminal A, (C) grade III Luminal A, (D) grade II LuminalB, (E) grade III Luminal B, (F) grade II Her2, (G) grade III Her2,(H) grade II triple negative, (I) grade III triple negative. Datashow specific staining patterns based upon grade and tissue subtype.

Mentions: A recentreport established that staining of histone H1 phosphorylationat T146 in bladder cancer tumor tissue can correlate with tumor grade.8 To establish the histone H1 pT146 staining patternsand proof of principle in vivo, we immunohistochemicallystained 242 primary tumor tissues taken from breast cancer patientswith carcinoma representing three tumor grades. Additionally, 97 nonbreastcancer tissues were stained for reference (Supplemental Data 10).Grade I tumor tissues are classified as well-differentiated, gradesII as intermediate, and grade III+ as poorly differentiated tissue.Figure 6A–C shows both the hematoxylinand eosin (H&E) and pT146 staining patterns across three gradesof Luminal A (estrogen receptor (ER)/progesterone receptor (PR) positive,human epithelial growth factor receptor (Her2) negative) primary tumors.The grade I tumor shows diffuse low intensity staining in the nucleiof tumor cells about the mammary ducts when stained for pT146 (Figure 6A). The grade II Luminal A tissues contain an overalllight speckled staining of the nuclei of tumor cells with individualvery intense stained nuclei scattered throughout the tissue (Figure 6B). The grade III tissues show a large number ofclustered and individual cells with intense nuclear pT146 staining(Figure 6C). The data show specific stainingpatterns across the three Luminal A breast tumor grades.


Histone H1 phosphorylation in breast cancer.

Harshman SW, Hoover ME, Huang C, Branson OE, Chaney SB, Cheney CM, Rosol TJ, Shapiro CL, Wysocki VH, Huebner K, Freitas MA - J. Proteome Res. (2014)

Immunohistochemicalstaining of primary breast tumorsfor Hematoxylin, eosin (H&E) and pT146: (A) grade I Luminal A,(B) grade II Luminal A, (C) grade III Luminal A, (D) grade II LuminalB, (E) grade III Luminal B, (F) grade II Her2, (G) grade III Her2,(H) grade II triple negative, (I) grade III triple negative. Datashow specific staining patterns based upon grade and tissue subtype.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4012839&req=5

fig6: Immunohistochemicalstaining of primary breast tumorsfor Hematoxylin, eosin (H&E) and pT146: (A) grade I Luminal A,(B) grade II Luminal A, (C) grade III Luminal A, (D) grade II LuminalB, (E) grade III Luminal B, (F) grade II Her2, (G) grade III Her2,(H) grade II triple negative, (I) grade III triple negative. Datashow specific staining patterns based upon grade and tissue subtype.
Mentions: A recentreport established that staining of histone H1 phosphorylationat T146 in bladder cancer tumor tissue can correlate with tumor grade.8 To establish the histone H1 pT146 staining patternsand proof of principle in vivo, we immunohistochemicallystained 242 primary tumor tissues taken from breast cancer patientswith carcinoma representing three tumor grades. Additionally, 97 nonbreastcancer tissues were stained for reference (Supplemental Data 10).Grade I tumor tissues are classified as well-differentiated, gradesII as intermediate, and grade III+ as poorly differentiated tissue.Figure 6A–C shows both the hematoxylinand eosin (H&E) and pT146 staining patterns across three gradesof Luminal A (estrogen receptor (ER)/progesterone receptor (PR) positive,human epithelial growth factor receptor (Her2) negative) primary tumors.The grade I tumor shows diffuse low intensity staining in the nucleiof tumor cells about the mammary ducts when stained for pT146 (Figure 6A). The grade II Luminal A tissues contain an overalllight speckled staining of the nuclei of tumor cells with individualvery intense stained nuclei scattered throughout the tissue (Figure 6B). The grade III tissues show a large number ofclustered and individual cells with intense nuclear pT146 staining(Figure 6C). The data show specific stainingpatterns across the three Luminal A breast tumor grades.

Bottom Line: The results show that the extent of H1 phosphorylation can distinguish between the different cell lines.The results suggest that the phosphorylation at threonine 146 is found on both histone H1.2 and histone H1.4.The data show that histone H1 phosphorylation can increase and decrease in response to extracellular stimuli.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Virology, Immunology and Medical Genetics, ‡Comprehensive Cancer Center, §Department of Chemistry & Biochemistry, ⊥Veterinary Biosciences, College of Veterinary Medicine, and ∥Department of Internal Medicine, Division of Hematology, The Ohio State University , Columbus, Ohio 43210, United States.

ABSTRACT
Breast cancer is the second leading cause of cancer-related deaths in women. The need for new clinical biomarkers in breast cancer is necessary to further predict prognosis and therapeutic response. In this article, the LC-MS histone H1 phosphorylation profiles were established for three distinct breast cancer cell lines. The results show that the extent of H1 phosphorylation can distinguish between the different cell lines. The histone H1 from the metastatic cell line, MDA-MB-231, was subjected to chemical derivitization and LC-MS/MS analysis. The results suggest that the phosphorylation at threonine 146 is found on both histone H1.2 and histone H1.4. Cell lines were then treated with an extracellular stimulus, estradiol or kinase inhibitor LY294002, to monitor changes in histone H1 phosphorylation. The data show that histone H1 phosphorylation can increase and decrease in response to extracellular stimuli. Finally, primary breast tissues were stained for the histone H1 phosphorylation at threonine 146. Variable staining patterns across tumor grades and subtypes were observed with pT146 labeling correlating with tumor grade. These results establish the potential for histone H1 phosphorylation at threonine 146 as a clinical biomarker in breast cancer.

Show MeSH
Related in: MedlinePlus