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Plasma Proteomic Profiling in Hereditary Breast Cancer Reveals a BRCA1-Specific Signature: Diagnostic and Functional Implications.

Scumaci D, Tammè L, Fiumara CV, Pappaianni G, Concolino A, Leone E, Faniello MC, Quaresima B, Ricevuto E, Costanzo FS, Cuda G - PLoS ONE (2015)

Bottom Line: Gene expression profiling studies have disclosed specific molecular signatures for BRCA1/2-related breast tumors as compared to sporadic cases, which might help diagnosis and clinical follow-up.To perform this analysis, we used samples from patients belonging to highly homogeneous population previously reported.The comparative analysis of the experimental results led to the identification of gelsolin as the most promising biomarker.

View Article: PubMed Central - PubMed

Affiliation: Dpt. of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta University Campus, Catanzaro, Italy.

ABSTRACT

Background: Breast cancer (BC) is a leading cause of death among women. Among the major risk factors, an important role is played by familial history of BC. Germ-line mutations in BRCA1/2 genes account for most of the hereditary breast and/or ovarian cancers. Gene expression profiling studies have disclosed specific molecular signatures for BRCA1/2-related breast tumors as compared to sporadic cases, which might help diagnosis and clinical follow-up. Even though, a clear hallmark of BRCA1/2-positive BC is still lacking. Many diseases are correlated with quantitative changes of proteins in body fluids. Plasma potentially carries important information whose knowledge could help to improve early disease detection, prognosis, and response to therapeutic treatments. The aim of this study was to develop a comprehensive approach finalized to improve the recovery of specific biomarkers from plasma samples of subjects affected by hereditary BC.

Methods: To perform this analysis, we used samples from patients belonging to highly homogeneous population previously reported. Depletion of high abundant plasma proteins, 2D gel analysis, liquid chromatography-tandem mass spectrometry (LC-MS/MS) and bioinformatics analysis were used into an integrated approach to investigate tumor-specific changes in the plasma proteome of BC patients and healthy family members sharing the same BRCA1 gene founder mutation (5083del19), previously reported by our group, with the aim to identify specific signatures.

Results: The comparative analysis of the experimental results led to the identification of gelsolin as the most promising biomarker.

Conclusions: Further analyses, performed using a panel of breast cancer cell lines, allowed us to further elucidate the signaling network that might modulate the expression of gelsolin in breast cancer.

No MeSH data available.


Related in: MedlinePlus

SDS-PAGE gels stained with Coomassie blue, before and after depletion of the high abundant proteins.Lanes 1, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 are crude plasma samples from subjects enrolled for the analysis. Lanes 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23 and 25 are MARS “Top-7” HPLC-treated plasma samples for depletion of the high abundance proteins. Lane 3 is empty; M: molecular weight standard. 20 μg of proteins were loaded in each lane.
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pone.0129762.g001: SDS-PAGE gels stained with Coomassie blue, before and after depletion of the high abundant proteins.Lanes 1, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 are crude plasma samples from subjects enrolled for the analysis. Lanes 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23 and 25 are MARS “Top-7” HPLC-treated plasma samples for depletion of the high abundance proteins. Lane 3 is empty; M: molecular weight standard. 20 μg of proteins were loaded in each lane.

Mentions: The large dynamic range of protein abundance in plasma represents a substantial analytical challenge. Removal of abundant plasma proteins using antibody capture approaches is a common and attractive mean to reduce sample complexity and to aid the analysis of lower abundance proteins of interest. Here, plasma most abundant proteins (albumin, IgG, antitrypsin, IgA, transferrin, haptoglobin, fibrinogen) were depleted using the MARS “Top-7” HPLC system (Agilent). HPLC depletion chromatogram for each run is shown in supporting S1 Fig. SDS-PAGE was applied to the samples before and after depletion to confirm the columns efficiency. As shown in Fig 1, the most abundant band around the 70 kDa area, which corresponds to albumin, was markedly reduced in the depleted samples. In addition, replicate analysis demonstrated high column reproducibility, linearity and efficient removal of abundant proteins.


Plasma Proteomic Profiling in Hereditary Breast Cancer Reveals a BRCA1-Specific Signature: Diagnostic and Functional Implications.

Scumaci D, Tammè L, Fiumara CV, Pappaianni G, Concolino A, Leone E, Faniello MC, Quaresima B, Ricevuto E, Costanzo FS, Cuda G - PLoS ONE (2015)

SDS-PAGE gels stained with Coomassie blue, before and after depletion of the high abundant proteins.Lanes 1, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 are crude plasma samples from subjects enrolled for the analysis. Lanes 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23 and 25 are MARS “Top-7” HPLC-treated plasma samples for depletion of the high abundance proteins. Lane 3 is empty; M: molecular weight standard. 20 μg of proteins were loaded in each lane.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465499&req=5

pone.0129762.g001: SDS-PAGE gels stained with Coomassie blue, before and after depletion of the high abundant proteins.Lanes 1, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 are crude plasma samples from subjects enrolled for the analysis. Lanes 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23 and 25 are MARS “Top-7” HPLC-treated plasma samples for depletion of the high abundance proteins. Lane 3 is empty; M: molecular weight standard. 20 μg of proteins were loaded in each lane.
Mentions: The large dynamic range of protein abundance in plasma represents a substantial analytical challenge. Removal of abundant plasma proteins using antibody capture approaches is a common and attractive mean to reduce sample complexity and to aid the analysis of lower abundance proteins of interest. Here, plasma most abundant proteins (albumin, IgG, antitrypsin, IgA, transferrin, haptoglobin, fibrinogen) were depleted using the MARS “Top-7” HPLC system (Agilent). HPLC depletion chromatogram for each run is shown in supporting S1 Fig. SDS-PAGE was applied to the samples before and after depletion to confirm the columns efficiency. As shown in Fig 1, the most abundant band around the 70 kDa area, which corresponds to albumin, was markedly reduced in the depleted samples. In addition, replicate analysis demonstrated high column reproducibility, linearity and efficient removal of abundant proteins.

Bottom Line: Gene expression profiling studies have disclosed specific molecular signatures for BRCA1/2-related breast tumors as compared to sporadic cases, which might help diagnosis and clinical follow-up.To perform this analysis, we used samples from patients belonging to highly homogeneous population previously reported.The comparative analysis of the experimental results led to the identification of gelsolin as the most promising biomarker.

View Article: PubMed Central - PubMed

Affiliation: Dpt. of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta University Campus, Catanzaro, Italy.

ABSTRACT

Background: Breast cancer (BC) is a leading cause of death among women. Among the major risk factors, an important role is played by familial history of BC. Germ-line mutations in BRCA1/2 genes account for most of the hereditary breast and/or ovarian cancers. Gene expression profiling studies have disclosed specific molecular signatures for BRCA1/2-related breast tumors as compared to sporadic cases, which might help diagnosis and clinical follow-up. Even though, a clear hallmark of BRCA1/2-positive BC is still lacking. Many diseases are correlated with quantitative changes of proteins in body fluids. Plasma potentially carries important information whose knowledge could help to improve early disease detection, prognosis, and response to therapeutic treatments. The aim of this study was to develop a comprehensive approach finalized to improve the recovery of specific biomarkers from plasma samples of subjects affected by hereditary BC.

Methods: To perform this analysis, we used samples from patients belonging to highly homogeneous population previously reported. Depletion of high abundant plasma proteins, 2D gel analysis, liquid chromatography-tandem mass spectrometry (LC-MS/MS) and bioinformatics analysis were used into an integrated approach to investigate tumor-specific changes in the plasma proteome of BC patients and healthy family members sharing the same BRCA1 gene founder mutation (5083del19), previously reported by our group, with the aim to identify specific signatures.

Results: The comparative analysis of the experimental results led to the identification of gelsolin as the most promising biomarker.

Conclusions: Further analyses, performed using a panel of breast cancer cell lines, allowed us to further elucidate the signaling network that might modulate the expression of gelsolin in breast cancer.

No MeSH data available.


Related in: MedlinePlus