Mass Spectrometry (LC–MS/MS) Identified Proteomic Biosignatures of Breast Cancer in Proximal Fluid

Whelan, Stephen A.; He, Jianbo; Lu, Ming; Souda, Puneet; Saxton, Romaine E.; Faull, Kym F.; Whitelegge, Julian P.; Chang, Helena R.

doi:10.1021/pr300606e

Cited by 44 publications

(42 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…aThe proteins that co-immunoprecipitated with NEK7-FLAG but not FLAG-control and that were not deposited in the CRAPome or the proteins that overlapped with Y2H are listed.bThe overlapping proteins found by mass spectrometry (this table) and by yeast two-hybrid (Table 1) are underlined.cFold change, as determined by the Scaffold program; 25 TIC = total ion count.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the Human NEK7 Interactome Suggests Catalytic and Regulatory Properties Distinct from Those of NEK6

et al. 2014

Self Cite

View full text Add to dashboard Cite

Human NEK7 is a regulator of cell division and plays an important role in growth and survival of mammalian cells. Human NEK6 and NEK7 are closely related, consisting of a conserved C-terminal catalytic domain and a nonconserved and disordered N-terminal regulatory domain, crucial to mediate the interactions with their respective proteins. Here, in order to better understand NEK7 cellular functions, we characterize the NEK7 interactome by two screening approaches: one using a yeast two-hybrid system and the other based on immunoprecipitation followed by mass spectrometry analysis. These approaches led to the identification of 61 NEK7 interactors that contribute to a variety of biological processes, including cell division. Combining additional interaction and phosphorylation assays from yeast two-hybrid screens, we validated CC2D1A, TUBB2B, MNAT1, and NEK9 proteins as potential NEK7 interactors and substrates. Notably, endogenous RGS2, TUBB, MNAT1, NEK9, and PLEKHA8 localized with NEK7 at key sites throughout the cell cycle, especially during mitosis and cytokinesis. Furthermore, we obtained evidence that the closely related kinases NEK6 and NEK7 do not share common interactors, with the exception of NEK9, and display different modes of protein interaction, depending on their N- and C-terminal regions, in distinct fashions. In summary, our work shows for the first time a comprehensive NEK7 interactome that, combined with functional in vitro and in vivo assays, suggests that NEK7 is a multifunctional kinase acting in different cellular processes in concert with cell division signaling and independently of NEK6.

show abstract

Section: Resultsmentioning

confidence: 99%

Characterization of the Human NEK7 Interactome Suggests Catalytic and Regulatory Properties Distinct from Those of NEK6

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…MCF-7 and DCIS.com cells, which are derived from non-metastatic breast cancers (67, 72), accumulate relatively low extracellular levels of LGALS3BP, while MDA-MB 231 cells, which are derived from a metastatic breast cancer (40), accumulate high extracellular levels of LGALS3BP (103–109). Patients with metastatic breast cancer tend to have abnormally high serum levels of LGALS3BP (50, 52).…”

Section: Discussionmentioning

confidence: 99%

Galectin-3 Binding Protein Secreted by Breast Cancer Cells Inhibits Monocyte-Derived Fibrocyte Differentiation

White

Roife

Gomer

2015

The Journal of Immunology

View full text Add to dashboard Cite

To metastasize, tumor cells often need to migrate through a layer of collagen-containing scar tissue which encapsulates the tumor. A key component of scar tissue and fibrosing diseases is the monocyte-derived fibrocyte, a collagen-secreting pro-fibrotic cell. To test the hypothesis that invasive tumor cells may block the formation of the fibrous sheath, we determined if tumor cells secrete factors that inhibit monocyte-derived fibrocyte differentiation. We found that the human metastatic breast cancer cell line MDA-MB 231 secretes activity that inhibits human monocyte-derived fibrocyte differentiation, while less aggressive breast cancer cell lines secrete less of this activity. Purification indicated that Galectin-3 Binding Protein (LGALS3BP) is the active factor. Recombinant LGALS3BP inhibits monocyte-derived fibrocyte differentiation, and immunodepletion of LGALS3BP from MDA-MB 231 conditioned media removes the monocyte-derived fibrocyte differentiation-inhibiting activity. LGALS3BP inhibits the differentiation of monocyte-derived fibrocytes from wild-type mouse spleen cells, but not from SIGN-R1−/− mouse spleen cells, suggesting that CD209/SIGN-R1 is required for the LGALS3BP effect. Galectin-3 and galectin-1, binding partners of LGALS3BP, potentiate monocyte-derived fibrocyte differentiation. In breast cancer biopsies, increased levels of tumor cell-associated LGALS3BP were observed in regions of the tumor that were invading the surrounding stroma. These findings suggest LGALS3BP and galectin-3 as new targets to treat metastatic cancer and fibrosing diseases.

show abstract

“…Comparative proteomic analysis of primary breast tumors and lymph node metastasis successfully identified recurrent differentially expressed proteins (Milioli et al, 2015). Additionally, proteomic-based research has brought new insights on therapy response (He et al, 2009), HER2 overexpression (Tang et al, 2013), and biomarker discovery in fluids (Whelan et al, 2012). In this report, we compare three IDC and three ILC samples using two-dimensional gel electrophoresis (2-DE) and MS to uncover specific protein biomarkers for each histological type.…”

Section: Introductionmentioning

confidence: 99%

Comparative proteomic analysis of ductal and lobular invasive breast carcinoma

Oliveira¹,

Gomig²,

Milioli³

et al. 2016

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Breast cancer is the second most common cancer worldwide and the first among women. Invasive ductal carcinoma (IDC) and invasive lobular carcinoma (ILC) are the two major histological subtypes, and the clinical and molecular differences between them justify the search for new markers to distinguish them. As proteomic analysis allows for a powerful and analytical approach to identify potential biomarkers, we performed a comparative analysis of IDC and ILC samples by using two-dimensional electrophoresis and mass spectrometry. Twenty-three spots were identified corresponding to 10 proteins differentially expressed between the two subtypes. ACTB, ACTG, TPM3, TBA1A, TBA1B, VIME, TPIS, PDIA3, PDIA6, and VTDB were upregulated in ductal carcinoma compared to in lobular carcinoma samples. Overall, these 10 proteins have a key role in oncogenesis. Their specific functions and relevance in cancer initiation and progression are further discussed in this study. The identified peptides represent promising biomarkers for the differentiation of ductal and lobular breast cancer subtypes, and for future interventions based on tailored therapy.

show abstract

Mass Spectrometry (LC–MS/MS) Identified Proteomic Biosignatures of Breast Cancer in Proximal Fluid

Cited by 44 publications

References 50 publications

Characterization of the Human NEK7 Interactome Suggests Catalytic and Regulatory Properties Distinct from Those of NEK6

Characterization of the Human NEK7 Interactome Suggests Catalytic and Regulatory Properties Distinct from Those of NEK6

Galectin-3 Binding Protein Secreted by Breast Cancer Cells Inhibits Monocyte-Derived Fibrocyte Differentiation

Comparative proteomic analysis of ductal and lobular invasive breast carcinoma

Contact Info

Product

Resources

About