Proteomic analyses of normal and malignant breast tissue—Identification of a tumor-specific protein expression profile

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 10524-10524
Author(s):  
G. Hudelist ◽  
C. Singer ◽  
K. Pischinger ◽  
K. Kaserer ◽  
M. Manavi ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Protein Expression ◽  
Antineoplastic Therapy ◽  
Proteomic Analysis ◽  
Protein Identification ◽  
Traditional Approach ◽  
Specific Protein ◽  
2D Page ◽  
Malignant Breast

10524 Background: Gene expression analysis has become a promising tool in predicting the clinical course of malignant disease and the response to antineoplastic therapy. Suprisingly, only little is known about the protein expression pattern of human tumors. Recent advances in proteomic analysis allow to identify proteins of interest by their expression and/or modification pattern in 2D-PAGE rather than using the traditional approach of translating gene expression data. Methods: In order to identify a proteomic pattern that is characteristic for malignant breast epithelium, we performed differential 2D-PAGE analysis in sets of microdissected malignant breast epithelia and corresponding adjacent normal breast epithelia from 5 patients with invasive breast carcinoma. Results: Thirty-two protein spots were found to be selectively regulated in malignant epithelium, and were subjected to MALDI-TOF and/or immunoblotting for protein identification. Thirteen of the identified proteins had previously not been associated with breast cancer. The validity of these findings was confirmed by literature review and immunohistochemistry for identified proteins in an independent cohort of 50 breast cancer specimens. Conclusions: We here describe a proteomic analysis of matched normal and malignant epithelia from invasive breast carcinomas. This strategy leads to a better understanding of oncogenesis at an operational level and helps to characterize the malignant phenotype of individual tumors and thereby to identify novel targets for antineoplastic therapy. No significant financial relationships to disclose.

Proteomic analysis in human breast cancer: Identification of a characteristic protein expression profile of malignant breast epithelium

PROTEOMICS ◽  
2006 ◽  
Vol 6 (6) ◽  
pp. 1989-2002 ◽  
Author(s):  
Gernot Hudelist ◽  
Christian F. Singer ◽  
Kerstin I. D. Pischinger ◽  
Klaus Kaserer ◽  
Mahmood Manavi ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Protein Expression ◽  
Expression Profile ◽  
Proteomic Analysis ◽  
Human Breast Cancer ◽  
Human Breast ◽  
Protein Expression Profile ◽  
Breast Epithelium ◽  
Malignant Breast

scholarly journals Protein citrullination as a source of cancer neoantigens

2021 ◽  
Vol 9 (6) ◽  
pp. e002549
Author(s):  
Hiroyuki Katayama ◽  
Makoto Kobayashi ◽  
Ehsan Irajizad ◽  
Alejandro Sevillarno ◽  
Nikul Patel ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Immune Response ◽  
Protein Expression ◽  
Cell Lines ◽  
Cancer Cell ◽  
Family Members ◽  
Immunohistochemical Analysis ◽  
Cancer Cell Lines ◽  
Mhc Ii

BackgroundCitrulline post-translational modification of proteins is mediated by protein arginine deiminase (PADI) family members and has been associated with autoimmune diseases. The role of PADI-citrullinome in immune response in cancer has not been evaluated. We hypothesized that PADI-mediated citrullinome is a source of neoantigens in cancer that induces immune response.MethodsProtein expression of PADI family members was evaluated in 196 cancer cell lines by means of indepth proteomic profiling. Gene expression was assessed using messenger RNA data sets from The Cancer Genome Atlas. Immunohistochemical analysis of PADI2 and peptidyl-citrulline was performed using breast cancer tissue sections. Citrullinated 12–34-mer peptides in the putative Major Histocompatibility Complex-II (MHC-II) binding range were profiled in breast cancer cell lines to investigate the relationship between protein citrullination and antigen presentation. We further evaluated immunoglobulin-bound citrullinome by mass spectrometry using 156 patients with breast cancer and 113 cancer-free controls.ResultsProteomic and gene expression analyses revealed PADI2 to be highly expressed in several cancer types including breast cancer. Immunohistochemical analysis of 422 breast tumor tissues revealed increased expression of PADI2 in ER− tumors (p<0.0001); PADI2 protein expression was positively correlated (p<0.0001) with peptidyl-citrulline staining. PADI2 expression exhibited strong positive correlations with a B cell immune signature and with MHC-II-bound citrullinated peptides. Increased circulating citrullinated antigen–antibody complexes occurred among newly diagnosed breast cancer cases relative to controls (p=0.0012).ConclusionsAn immune response associated with citrullinome is a rich source of neoantigens in breast cancer with a potential for diagnostic and therapeutic applications.

scholarly journals 83 Spatially resolved transcriptomic and proteomic investigation of breast cancer and its immune microenvironment

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A91-A91
Author(s):  
Jennifer Chew ◽  
Cedric Uytingco ◽  
Rapolas Spalinskas ◽  
Yifeng Yin ◽  
Joe Shuga ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Tumor Microenvironment ◽  
Protein Expression ◽  
Protein Detection ◽  
Response To Therapy ◽  
Pathological Examination ◽  
Multi Drug Resistance ◽  
Spatially Resolved ◽  
Gene And Protein Expression

BackgroundThe tumor microenvironment (TME) is composed of highly heterogeneous extracellular structures and cell types such as endothelial cells, immune cells, and fibroblasts that dynamically influence and communicate with each other. The constant interaction between a tumor and its microenvironment plays a critical role in cancer development and progression and can significantly affect a tumor’s response to therapy and capacity for multi-drug resistance. High resolution analyses of gene and protein expression with spatial context can provide deeper insights into the interactions between tumor cells and surrounding cells within the TME, where a better understanding of the underlying biology can improve treatment efficacy and patient outcomes. Here, we demonstrated the ability to perform streamlined multi-omic tumor analyses by utilizing the 10X Genomics Visium Spatial Gene Expression Solution for FFPE with multiplex protein enablement. This technique simultaneously assesses gene and protein expression to elucidate the immunological profile and microenvironment of different breast cancer samples in conjunction with standard pathological methods.MethodsSerial (5 µm) sections of FFPE human breast cancer samples were placed on Visium Gene Expression (GEX) slides. The Visium GEX slides incorporate ~5,000 molecularly barcoded, spatially encoded capture spots onto which tissue sections are placed, stained, and imaged. Following incubation with a human whole transcriptome, probe-based RNA panel and an immuno-oncology oligo-tagged antibody panel, developed with Abcam conjugated antibodies, the tissues are permeabilized and the representative probes are captured. Paired GEX and protein libraries are generated for each section and then sequenced on an Illumina NovaSeq at a depth of ~50,000 reads per spot. Resulting reads from both libraries are aligned and overlaid with H&E-stained tissue images, enabling analysis of both mRNA and protein expression. Additional analyses and data visualizations were performed on the Loupe Browser v4.1 desktop software.ConclusionsSpatial transcriptomics technology complements pathological examination by combining histological assessment with the throughput and deep biological insight of highly-multiplexed protein detection and RNA-seq. Taken together, our work demonstrated that Visium Spatial technology provides a spatially-resolved, multi-analyte view of the tumor microenvironment, where a greater understanding of cellular behavior in and around tumors can help drive discovery of new biomarkers and therapeutic targets.

Effect of Auraptene on angiogenesis in Xenograft model of breast cancer

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Mohammad Reza Shiran ◽  
Elham Mahmoudian ◽  
Abolghasem Ajami ◽  
Seyed Mostafa Hosseini ◽  
Ayjamal Khojasteh ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Animal Model ◽  
Protein Expression ◽  
Western Blot ◽  
Xenograft Model ◽  
Dependent Manner ◽  
Concentration Dependent Manner

Abstract Objectives Angiogenesis is the most important challenge in breast cancer treatment. Recently, scientists become interesting in rare natural products and intensive researches was performed to identify their pharmacological profile. Auraptene shows helpful effects such as cancer chemo-preventive, anti-inflammatory, anti-oxidant, immuno-modulatory. In this regard, we investigated the anti-angiogenesis effect of Auraptene in in-vitro and in-vivo model of breast cancer. Methods In this study, 4T, MDA-MB-231 and HUVEC cell lines were used. The proliferation study was done by MTT assay. For tube formation assay, 250 matrigel, 1 × 104 HUVEC treated with Auraptene, 20 ng/mL EGF, 20 ng/mL bFGF and 20 ng/mL VEGF were used. Gene expression of important gene related to angiogenesis in animal model of breast cancer was investigated by Real-time PCR. Protein expression of VCAM-1 and TNFR-1 gene related to angiogenesis in animal model of breast cancer was investigated by western-blot. Results Auraptene treatment led to reduction in cell viability of MDA-MB-231 in a concentration-dependent manner. Also, we observed change in the number of tubes or branches formed by cells incubated with 40 and 80 μM Auraptene. Auraptene effect the gene expression of important gene related to angiogenesis (VEGF, VEGFR2, COX2, IFNɣ). Moreover, the western blot data exhibited that Auraptene effect the protein expression of VCAM-1 and TNFR-1. Conclusions Overall, this study shows that Auraptene significantly suppressed angiogenesis via down-regulation of VEGF, VEGFR2, VCAM-1, TNFR-1, COX-2 and up-regulation of IFNγ.

scholarly journals RNA interference in gene cloning/expression

2016 ◽  
Author(s):  
Vikash Bhardwaj
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Protein Expression ◽  
Gene Cloning ◽  
Recombinant Plasmid ◽  
Specific Protein ◽  
Dna Cloning ◽  
Production Stage ◽  
False Conclusion ◽  
Or Gene

DNA cloning is a simple, straightforward process. It has been put into practice as a standard lab procedure for cloning the gene of interest for specific protein expression and other molecular or cellular studies. The present study is the first of its kind to hypothesize the formation of interfering recombinant plasmid during a DNA cloning reaction that is believed to interfere with the protein or gene expression. This might lead to a false conclusion in a cellular study and also may lower the protein expression during the production stage.

Response of angiogenic factors to the treatment with melatonin in breast cancer cell lines.

2012 ◽  
Vol 30 (27_suppl) ◽  
pp. 120-120
Author(s):  
Bruna Victorasso Jardim ◽  
Marina Gobbe Moschetta ◽  
Thaiz Ferraz Borin ◽  
Gabriela Bottaro Gelaleti ◽  
Camila Leonel ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Protein Expression ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Therapeutic Agent ◽  
Breast Cancer Cell Lines ◽  
Er Positive ◽  
Vegf Protein ◽  
Mcf 7

120 Background: Tumor growth and metastasis are the leading cause of death in breast cancer patients. The tumor growth requires neoangiogenesis, stimulated by vascular endothelial growth factor (VEGF) expressed under control of hypoxia-inducible factor-1α (HIF-1α). Melatonin, a natural hormone has been suggested as a new therapeutic agent to inhibit angiogenesis factors in several types of cancers. We evaluated the HIF-1α and VEGF expression in breast cancer cell line after hypoxia and treatment with melatonin. Methods: Breast cancer cell lines ER-positive (MCF-7) and ER-negative (MDA-MB-231) were cultured in DMEM at 37°C in 5% CO2. The cells were treated with CoCl2 to mimic hypoxia, and then treated with melatonin. Cell viability was measured by MTT assay with five melatonin concentrations (0.5mM, 1mM, 2mM, 5mM, 10mM). For the pharmacological concentration of melatonin (1mM) the protein and gene expression of HIF-1α and VEGF were detected by immunocytochemistry and real time PCR, respectively. Results: There was a significantly decrease in MCF-7 cells viability after treatment with all concentrations of melatonin and in MDA-MB-231 cells just with 1mM (p<0.05). The melatonin treatment did not alter the protein and gene expression of HIF-1α in MCF-7 cells (p>0.05), however, it significantly decreased the VEGF protein expression (p<0.05). In MDA-MB-231 cells there was significant decreased in the HIF-1α protein expression (p<0.05). Conclusions: Our results showed that 1mM of melatonin decreased the cell viability and HIF-1α protein expression in ER-negative cells, and VEGF protein expression in ER-positive cells. This study is the first that evaluated the expression of these angiogenic factors in breast cancer cells after treatment with melatonin that can be a potential therapeutic agent. Thus, we are performing in vivo study to confirm the potential effectiveness of melatonin in the control of angiogenesis in breast cancer.

Proteomic Analysis of the Specific Protein Expression in Nasal Polyp

2009 ◽  
Vol 52 (3) ◽  
pp. 228
Author(s):  
Song-Chul Park ◽  
Seung Hyun Koh ◽  
Sang Heon Lee ◽  
Jae Hoon Lee
Keyword(s):  
Protein Expression ◽  
Nasal Polyp ◽  
Proteomic Analysis ◽  
Specific Protein

Comprehensive Analysis of the Expression and Prognostic Significance of THBSs in Breast Cancer

2021 ◽  
Author(s):  
Jun Wang ◽  
Xuebing Zhan ◽  
Qian Luo ◽  
Yunshu Kuang ◽  
Xiao Liang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Protein Expression ◽  
Cancer Patients ◽  
Prognostic Value ◽  
Breast Cancer Patients ◽  
Free Survival ◽  
Expression Levels ◽  
Cancer Tissues ◽  
Mrna Expression Levels

Abstract Background: Breast cancer is one of the most common tumors for women worldwide. Thrombospondins (THBSs) are reported to play important roles in various cellular processes and are involved in the occurrence and development of human cancers. However, the expression and prognostic value of THBSs family in breast cancer remain unclear.Methods: In this study, we examined the genes and protein expression levels of THBSs and their prognostic value by synthesizing several mainstream databases, including Oncomine, Human Protein Atlas (HPA), UALCAN, and KM Plotter. We also analyzed THBS interaction networks, genetic alterations, functional enrichment, and drug sensitivity with several publicly accessible databases, including GEPIA, GeneMANIA, STRING, cBioPortal, Metascape and NCI-60 database.Results: The results showed that the mRNA expression levels of THBS1, THBS2, THBS3, and THBS5 in breast cancer tissues were significantly higher than in normal tissues. The mRNA expression levels of THBS4 were different in different subtypes of breast cancer, and the protein expression levels of THBS1, THBS2, and THBS4 in breast cancer tissues were higher than in normal breast tissues. Survival analysis showed that breast cancer patients with high THBS1 gene expression showed worse overall survival (OS), relapse-free survival (RFS), and post-progression survival (PPS), and breast cancer patients with high THBS2 gene expression also showed worse RFS. Conversely, lower THBS3 levels predicted worse RFS, and lower THBS4 levels predicted worse OS, RFS, and distant metastasis-free survival (DMFS). Conclusions: These results suggest that THBSs may be potential biomarkers for breast cancer.

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