Identification of Candidate Biomarker Proteins Released by Human Endometrial and Cervical Cancer Cells Using Two-Dimensional Liquid Chromatography/Tandem Mass Spectrometry

Li, Hongyan; DeSouza, Leroi V.; Ghanny, Shaun; Li, Wei; Romaschin, Alexander D.; Colgan, Terence J.; Siu, Kin Wai Michael

doi:10.1021/pr0700798

Cited by 27 publications

(19 citation statements)

References 72 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A total of 56 CD antigens were detected among the HeLa cell proteins, accounting for nearly 25% of the glycoproteins identified (supplemental Table 2). The effectiveness of surface membrane enrichment from our results is comparable with other reported studies (38,(41)(42)(43)(44)(45). We also detected other classes of surface membrane proteins including GPI-anchored proteins.…”

Section: N-linked Glycosylation Sites Identified In the M Amurensis supporting

confidence: 91%

Combining Results from Lectin Affinity Chromatography and Glycocapture Approaches Substantially Improves the Coverage of the Glycoproteome

McDonald

Yang

Marathe

et al. 2009

Molecular & Cellular Proteomics

110

112

View full text Add to dashboard Cite

Identification of glycosylated proteins, especially those in the plasma membrane, has the potential of defining diagnostic biomarkers and therapeutic targets as well as increasing our understanding of changes occurring in the glycoproteome during normal differentiation and disease processes. Although many cellular proteins are glycosylated they are rarely identified by mass spectrometric analysis (e.g. shotgun proteomics) of total cell lysates. Therefore, methods that specifically target glycoproteins are necessary to facilitate their isolation from total cell lysates prior to their identification by mass spectrometrybased analysis. To enrich for plasma membrane glycoproteins the methods must selectively target characteristics associated with proteins within this compartment. We demonstrate that the application of two methods, one that uses periodate to label glycoproteins of intact cells and a hydrazide resin to capture the labeled glycoproteins and another that targets glycoproteins with sialic acid residues using lectin affinity chromatography, in conjunction with liquid chromatography-tandem mass spectrometry is effective for plasma membrane glycoprotein identification. We demonstrate that this combination of methods dramatically increases coverage of the plasma membrane proteome (more than one-half of the membrane glycoproteins were identified by the two methods uniquely) and also results in the identification of a large number of secreted glycoproteins. Our approach avoids the need for subcellular fractionation and utilizes a simple detergent lysis step that effectively solubilizes membrane glycopro-

show abstract

Section: N-linked Glycosylation Sites Identified In the M Amurensis supporting

confidence: 91%

Combining Results from Lectin Affinity Chromatography and Glycocapture Approaches Substantially Improves the Coverage of the Glycoproteome

McDonald

Yang

Marathe

et al. 2009

Molecular & Cellular Proteomics

110

112

View full text Add to dashboard Cite

show abstract

“…[34][35][36][37] As a result, the screening for disease markers became one of the main topics in a large number of proteomic studies in the past decade. [38][39][40][41][42] In this study, we performed a proteomic analysis to investigate tumor-specific protein expression of human breast carcinoma tissues. Our results showed that proteins involved in antiapoptosis, cell motility, cell proliferation, glycolysis, protein folding, signal transduction, and other processes related to tumorigenesis were overexpressed in cancer tissues.…”

Section: Discussionmentioning

confidence: 99%

Targeting Therapy for Breast Carcinoma by ATP Synthase Inhibitor Aurovertin B

et al. 2008

View full text Add to dashboard Cite

Targeting of tumor tissues is one of the most powerful approaches to accelerate the efficiency of anticancer treatments. The investigation of effective targets, including proteins specifically and abundantly expressed in abnormal regions, has been one of the most important research topics in cancer therapy. In this study, we performed a proteomic analysis on human breast carcinoma tissues to investigate the tumor-specific protein expression in breast carcinoma. Our study showed that ATP synthase was up-regulated in tumor tissues and was present on the plasma membrane of breast cancer cells. Furthermore, we treated the breast cancer cells with ATP synthase inhibitors and examined the inhibitory efficiency. Aurovertin B, an ATP synthase inhibitor, has strong inhibition on the proliferation of several breast cancer cell lines, but little influence on the normal cell line MCF-10A. Aurovertin B inhibits proliferation of breast cancer cells by inducing apoptosis and arresting cell cycle at the G0/G1 phase. This study showed aurovertin B can be used as an antitumorigenic agent and may be exploited in cancer chemotherapy.

show abstract

“…Acetone-precipitated GBM secretome proteins (50 lg) were digested with trypsin (Promega) using an established in-solution digestion protocol [13]. Peptides were separated by two-dimensional liquid chromatography with strong cation exchange being the first dimension, followed by reverse phase chromatography in the second dimension as previously published [14].…”

Section: Mass Spectrometry Analysis Of Gbm Secretomementioning

confidence: 99%

GBM secretome induces transient transformation of human neural precursor cells

et al. 2012

Self Cite

View full text Add to dashboard Cite

Glioblastoma (GBM) is the most aggressive primary brain tumor in humans, with a uniformly poor prognosis. The tumor microenvironment is composed of both supportive cellular substrates and exogenous factors. We hypothesize that exogenous factors secreted by brain tumor initiating cells (BTICs) could predispose normal neural precursor cells (NPCs) to transformation. When NPCs are grown in GBM-conditioned media, and designated as "tumor-conditioned NPCs" (tcNPCs), they become highly proliferative and exhibit increased stem cell self-renewal, or the unique ability of stem cells to asymmetrically generate another stem cell and a daughter cell. tcNPCs also show an increased transcript level of stem cell markers such as CD133 and ALDH and growth factor receptors such as VEGFR1, VEGFR2, EGFR and PDGFRα. Media analysis by ELISA of GBM-conditioned media reveals an elevated secretion of growth factors such as EGF, VEGF and PDGF-AA when compared to normal neural stem cell-conditioned media. We also demonstrate that tcNPCs require prolonged or continuous exposure to the GBM secretome in vitro to retain GBM BTIC characteristics. Our in vivo studies reveal that tcNPCs are unable to form tumors, confirming that irreversible transformation events may require sustained or prolonged presence of the GBM secretome. Analysis of GBM-conditioned media by mass spectrometry reveals the presence of secreted proteins Chitinase-3-like 1 (CHI3L1) and H2A histone family member H2AX. Collectively, our data suggest that GBM-secreted factors are capable of transiently altering normal NPCs, although for retention of the transformed phenotype, sustained or prolonged secretome exposure or additional transformation events are likely necessary.

show abstract

Identification of Candidate Biomarker Proteins Released by Human Endometrial and Cervical Cancer Cells Using Two-Dimensional Liquid Chromatography/Tandem Mass Spectrometry

Abstract: Candidate biomarker proteins, including chaperonin 10 and pyruvate kinase, previously discovered and identified using mass-tagging reagents with multidimensional liquid chromatography and tandem mass spectrometry (

Cited by 27 publications

References 72 publications

Combining Results from Lectin Affinity Chromatography and Glycocapture Approaches Substantially Improves the Coverage of the Glycoproteome

Combining Results from Lectin Affinity Chromatography and Glycocapture Approaches Substantially Improves the Coverage of the Glycoproteome

Targeting Therapy for Breast Carcinoma by ATP Synthase Inhibitor Aurovertin B

GBM secretome induces transient transformation of human neural precursor cells

Contact Info

Product

Resources

About