Protein profiling of angiogenesis-related growth factors in laryngeal carcinoma: Pattern of protein expression in relation to tumour progression

Abstract Heterogeneity of glioblastoma (GBM) has been extensively studied in recent years with identification of oncogenic drivers of GBM cellular subtypes. However, little is known about how these cells interact with each other or with the surrounding tumor microenvironment (TME). We employed spatial protein profiling targeting immune and neuronal markers (79 proteins) coupled with single-cell spatial maps of fluorescence in situ hybridization (FISH) for EGFR, CDK4, and PDGFRA on human GBM tissue sections. Several cores from 20 GBM samples were collected to create a tissue microarray, and 96 regions of interests were profiled with 37,844 nuclei for oncogenic amplification screen. Spatial protein profiling identified strong correlation of certain immune markers, TAU-associated proteins, and oligodendrocyte-enriched protein groups and overall high intratumor heterogeneity of TME. Our single-cell quantification of FISH signals showed differences among tumors based on the prevalence of dual amplification of EGFR and CDK4 within a cell relative to single oncogene amplified cells. High relative frequency of dual amplification was associated with increased expression of immune-related markers and decreased expression of EGFR protein. Moreover, this protein expression signature was associated with survival in another GBM dataset. Here, we present spatial genetic analysis at the single cell level coupled with protein expression profiles associated with tumor microenvironment. Our results suggest that assessment of genetic heterogeneity in GBM could potentially drive improved patient stratification and treatment.

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Diagnostic impact of promoter methylation and E-cadherin gene and protein expression levels in laryngeal carcinoma

Współczesna Onkologia ◽

10.5114/wo.2013.35284 ◽

2013 ◽

Vol 3 ◽

pp. 263-271

Author(s):

Katarzyna Starska ◽

Ewa Forma ◽

Iwona Lewy-Trenda ◽

Paweł Papież ◽

Jan Woś ◽

...

Keyword(s):

Protein Expression ◽

Promoter Methylation ◽

Laryngeal Carcinoma ◽

Expression Levels ◽

Diagnostic Impact ◽

Gene And Protein Expression ◽

E Cadherin

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Molecular differences between normal breast epithelia, pure DCIS, and DCIS adjacent to invasion.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.27_suppl.50 ◽

2012 ◽

Vol 30 (27_suppl) ◽

pp. 50-50

Author(s):

Marina Guvakova

Keyword(s):

Breast Cancer ◽

Protein Expression ◽

Cancer Progression ◽

Rho Gtpase ◽

Ductal Carcinoma ◽

Continuous Variable ◽

Protein Profiling ◽

Nucleotide Exchange ◽

Fixed Tissue ◽

Gene And Protein Expression

50 Background: With the use of screening mammography, the diagnosis of breast ductal carcinoma in situ (DCIS) is increasing worldwide. Currently there are no applied molecular markers to aid in predicting risk of DCIS progression to invasive breast cancer (IBC). Studies predicted that the transition from DCIS to invasive disease was associated with quantitative rather than qualitative differences in gene and protein expression. Methods: We developed monochrome imaging-based method to measure protein expression as a continuous variable in fixed tissue (Furstenau et al. BREA 2010). The insulin-like growth factor I receptor (IGF-IR), Ras oncogene –like protein 1 (Rap1), and a Rho GTPase guanine nucleotide exchange factor Vav2 implicated in the regulation of invasion in preclinical models have been chosen for protein analysis in tissue. In pilot study, we performed quantitative protein profiling on 90 samples of the breast: 17 histologically normal tissues, 16 benign lesions; 15 CIS, and 42 IBC. In addition, we analyzed 24 DCIS: 12 pure and 12 associated with IBC. Results: The expression of the IGF-IR and Rap1 was increased in pure CIS; significantly (P ≤ 0.001) increased in CIS adjacent to IBC as well as in IBC compared with non-cancerous tissue. In the majority of pure DCIS the levels of Vav2 were similar to that in normal epithelia; however, the Vav2 levels were significantly (P<0.01) higher in DCIS adjacent to invasion and in IBC. Moreover, IGF-IR, Rap1 and Vav2 significantly discriminated invasive from non-invasive tissues, with receiver operating characteristic (ROC) area under curve, AUC=0.8. Interestingly, younger women (< 50 years of age) were more likely to develop IBC with increased expression of the Vav2 protein (OR= 2.05; 95% CI 1.08-3.86). No significant correlation was found between IGF-IR, Rap1 expression levels and patient’s age. Conclusions: We conclude that our assessment of protein expression in breast tissue revealed previously unidentified quantitative differences in the IGF-IR, Rap1, and Vav2 protein expression in breast cancer progression series. These findings open door for studies on molecular-based prediction of individualized risk for developing invasion in early diagnosed DCIS.

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The short and the long: non-coding RNAs and growth factors in cancer progression

Biochemical Society Transactions ◽

10.1042/bst20160131 ◽

2017 ◽

Vol 45 (1) ◽

pp. 51-64 ◽

Cited By ~ 16

Author(s):

Aldema Sas-Chen ◽

Swati Srivastava ◽

Yosef Yarden

Keyword(s):

Growth Factors ◽

Cancer Progression ◽

Tumour Progression ◽

Therapeutic Strategies ◽

Signalling Pathways ◽

Circulation System ◽

Multistep Process ◽

New Locations ◽

Non Coding Rnas ◽

Lines Of Evidence

A relatively well-understood multistep process enables mutation-bearing cells to form primary tumours, which later use the circulation system to colonize new locations and form metastases. However, in which way the emerging abundance of different non-coding RNAs supports tumour progression is poorly understood. Here, we review new lines of evidence linking long and short types of non-coding RNAs to signalling pathways activated in the course of cancer progression by growth factors and by the tumour micro-environment. Resolving the new dimension of non-coding RNAs in oncogenesis will probably translate to earlier detection of cancer and improved therapeutic strategies.

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MALDI-TOF mass spectrometric protein profiling of THP-1 cells and their microvesicles

Medical Immunology (Russia) ◽

10.15789/1563-0625-mtm-2141 ◽

2021 ◽

Vol 23 (2) ◽

pp. 275-292

Author(s):

A. V. Korenevsky ◽

Yu. P. Milyutina ◽

M. E. Berezkina ◽

E. P. Alexandrova ◽

O. A. Balabas ◽

...

Keyword(s):

Immune Response ◽

Growth Factors ◽

Cell Differentiation ◽

Functional Groups ◽

Extracellular Vesicles ◽

Intracellular Signaling ◽

Plasma Membranes ◽

Mass Spectrometric ◽

Protein Profiling ◽

Wide Range

Extracellular vesicles that are shed from the plasma membranes take an active part in intercellular communication, transporting a wide range of molecules, including proteins, lipids, nucleic acids and carbohydrates, being of great functional importance. One of the steps to better understanding of distant communications of cells and their regulatory mechanisms is a proteomic study of various extracellular vesicles, including microvesicles and exosomes. Pro-inflammatory cytokines produced by monocytes and individual complement system components play a key role in their specific functioning. The aim of this work was to study proteomic composition of THP-1 monocyte-like cells and their microvesicles. The MALDI-mass spectrometric analysis of electrophoretic protein fractions of cell lysates and microvesicles allowed for identifying 107 proteins that perform various functions. Among 19 determined functional groups, the largest ones comprise transcription regulators and proteins with unknown functions. The smallest functional groups include regulators of cell differentiation and development, proteins participating in immune response and inflammation, cellular receptors and their regulators, transporter and transport regulatory proteins, as well as cell proteins mediating adhesion and matrix structures, processing regulators, proteins of ubiquitin-proteasome system, intracellular signaling, autophagy and exocytosis regulators, chromatin structural proteins, hemostatic regulators, and peptide hormones. An intermediate position is occupied by cytokines and growth factors, enzymes, cytoskeleton and motor proteins, as well as RNA processing and translation regulators. The subsequent DAVID Functional Annotation Clustering analysis allowed for identifying the most common groups distributed by their molecular function, biological processes, and cellular component. Separately, in the microvesicles derived from THP-1 monocyte-like cells, proteins of the immune response and inflammation, cytokines and growth factors, intracellular signaling proteins, cell differentiation regulators and developmental proteins, as well as cell adhesion and matrix proteins were identified among other protein molecules. The data obtained on the partial proteome of THP-1 monocyte-like cells and their microvesicles extend the existing knowledge on distant communications between the cells and suggest new mechanisms of interaction between monocytes/macrophages and their microenvironment.

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CHITINASE-LIKE PROTEINS AS PROMISING MARKERS IN CANCER PATIENTS

Siberian Journal of Oncology ◽

10.21294/1814-4861-2018-17-4-99-105 ◽

2018 ◽

Vol 17 (4) ◽

pp. 99-105 ◽

Cited By ~ 2

Author(s):

I. V. Larionova ◽

T. N. Sevastyanova ◽

A. A. Rakina ◽

N. V. Cherdyntseva ◽

Ju. G. Kzhyshkowska

Keyword(s):

Growth Factors ◽

Cancer Patient ◽

Cancer Patients ◽

Inflammatory Diseases ◽

Tumour Progression ◽

Patient Benefit ◽

Pathological Conditions ◽

The Family ◽

Metastasis Development

In the present review we collected the main studies regarding the role of chitinase-like proteins (CLPs), belonging to the family of Glyco_18 domain-containing proteins, in different cancers. In humans, 3 chitinaselike proteins have been identified: YKL-40 (CHI3L1), YKL-39 (CHI3L2) and stabilin-1-interacting chitinase-like protein (SI-CLP). CLPs are produced by several types of cells and combine the properties of cytokines and growth factors. The high levels of CLPs were identified in the circulation of the patients with inflammatory diseases and various types of tumors. We highlighted the main known functions of CLPs in normal and pathological conditions, their contribution to metastasis development, angiogenesis, invasion and other processes in cancer, the correlation of the levels of CLPs with tumour progression. Our data also contribute to the understanding of question how CLP could be useful for cancer patient benefit.

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Oncosuppressive Role of RUNX3 in Human Astrocytomas

Journal of Oncology ◽

10.1155/2019/1232434 ◽

2019 ◽

Vol 2019 ◽

pp. 1-10

Author(s):

Giedrius Steponaitis ◽

Arunas Kazlauskas ◽

Paulina Vaitkienė ◽

Vytenis P. Deltuva ◽

Mykolas Mikuciunas ◽

...

Keyword(s):

Protein Expression ◽

Protein Level ◽

Tumour Progression ◽

Methylation Status ◽

Suppressor Gene ◽

Tumour Suppressor Gene ◽

Molecular Profile ◽

Prognostic Information ◽

Specific Pcr ◽

Related Transcription Factor

Background. Gliomas are the most common and aggressive among primary malignant brain tumours with significant inter- and intratumour heterogeneity in histology, molecular profile, and patient outcome. However, molecular targets that could provide reliable diagnostic and prognostic information on this type of cancer are currently unknown. Recent studies show that certain phenotypes of gliomas such as malignancy, resistance to therapy, and relapses are associated with the epigenetic alterations of tumour-specific genes. Runt-related transcription factor 3 (RUNX3) is feasible tumour suppressor gene since its inactivation was shown to be related to carcinogenesis. Aim. The aim of the study was to elucidate RUNX3 changes in different regulation levels of molecular biology starting from epigenetics to function in particular cases of astrocytic origin tumours of different grade evaluating significance of molecular changes of RUNX3 for patient clinical characteristics as well as evaluate RUNX3 reexpression effect to GBM cells. Methods. The methylation status and protein expression levels of RUNX3 were measured by methylation-specific PCR and Western blot in 136 and 72 different malignancy grade glioma tissues, respectively. Lipotransfection and MTT were applied for proliferation assessment in U87-MG cells. Results. We found that RUNX3 was highly methylated and downregulated in GBM. RUNX3 promoter methylation was detected in 69.4% of GBM (n=49) as compared to 0 to 17.2% in I-III grade astrocytomas (n=87). Weighty lower RUNX3 protein level was observed in GMB specimens compared to grade II-III astrocytomas. Correlation test revealed a weak but significant link among Runx3 methylation and protein level. Kaplan-Meier analysis showed that increased RUNX3 methylation and low protein level were both associated with shorter patient survival (p<0.05). Reexpression of RUNX3 in U87-MG cells significantly reduced glioma cell viability compared to control transfection. Conclusions. The results demonstrate that RUNX3 gene methylation and protein expression downregulation are glioma malignancy dependent and contribute to tumour progression.

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