scholarly journals Protocol for PTEN Expression by Immunohistochemistry in Formalin-fixed Paraffin-embedded Human Breast Carcinoma

2010 ◽  
Vol 18 (4) ◽  
pp. 371-374 ◽  
Author(s):  
Rita A. Sakr ◽  
Violetta Barbashina ◽  
Mary Morrogh ◽  
Sarat Chandarlapaty ◽  
Victor P. Andrade ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Human Breast Carcinoma ◽  
Pten Expression ◽  
Human Breast ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Expression of proliferation genes in formalin-fixed paraffin-embedded (FFPE) tissue from breast carcinomas. Feasibility and relevance for a routine histopathology laboratory

2016 ◽  
Vol 70 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Carla Thomas ◽  
Cleo Robinson ◽  
Ben Dessauvagie ◽  
Benjamin Wood ◽  
Greg Sterrett ◽  
...  
Keyword(s):  
Gene Expression ◽  
Breast Carcinoma ◽  
Expression Profiling ◽  
Proliferative Activity ◽  
Breast Cancers ◽  
Breast Carcinomas ◽  
Ki 67 ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

AimBreast carcinoma proliferative activity, histological grade and commercial molecular tests are all important in prognostication and treatment. There is a particular need for improved, standardised techniques for subclassification of grade 2 breast cancers into low-risk and high-risk prognostic groups. In this study we investigated whether gene expression profiling of five proliferation genes was feasible using breast cancer tissue in a clinical setting and whether these profiles could enhance pathological assessment.MethodsExpression of five proliferation gene mRNAs; Ki-67, STK 15, CCNB1, CCND1 and MYBL2, was quantified in 27 breast carcinomas and compared with Ki-67 proliferation index (PI) and Nottingham mitotic score.ResultsExpression of Ki-67, STK15 and MYBL2 mRNA showed moderate Spearman's correlation with Ki-67 PI (p<0.01), but CCND1 and CCNB1 showed weak, non-significant correlation. Individual gene expression did not associate with mitotic score but combined mRNA expression correlated with both Ki-67 PI (p=0.018) and mitotic score (p=0.03; 0.007).ConclusionsThis study confirms mRNA analysis in breast carcinoma formalin-fixed, paraffin-embedded samples is feasible and suggests gene expression profiling, using a small set of five proliferation genes, has potential in aiding histological grading or assessment of proliferative activity of breast cancers. To fully evaluate the clinical applicability of this approach, a larger cohort study with long-term follow-up data is required.

scholarly journals Single-day HER2neu amplification assessment using chip-based digital PCR in formalin-fixed paraffin-embedded breast carcinoma tissue

2018 ◽  
Vol Volume 10 ◽  
pp. 121-129
Author(s):  
Parth Shah ◽  
Shiva Murarka ◽  
Anupam Joshi ◽  
Bhavana Mehta ◽  
Vipal Parmar ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Digital Pcr ◽  
Carcinoma Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

scholarly journals Frequency of Epstein–Barr Virus DNA in Formalin-fixed Paraffin-embedded Tissue of Patients with Ductal Breast Carcinoma

2019 ◽  
Vol 20 (3) ◽  
pp. 687-692 ◽  
Author(s):  
Chia Sharifpour ◽  
Manoochehr Makvandi ◽  
Alireza Samabafzadeh ◽  
Abdolhassan Talaei-Zadeh ◽  
Nastaran Ranjbari ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Epstein Barr Virus ◽  
Barr Virus ◽  
Ductal Breast Carcinoma ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Epstein Barr ◽  
Formalin Fixed

scholarly journals Droplet Digital PCR Measurement of HER2 Copy Number Alteration in Formalin-Fixed Paraffin-Embedded Breast Carcinoma Tissue

2013 ◽  
Vol 59 (6) ◽  
pp. 991-994 ◽  
Author(s):  
Phillip Belgrader ◽  
Stephanie C Tanner ◽  
John F Regan ◽  
Ryan Koehler ◽  
Benjamin J Hindson ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Copy Number ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Invasive Breast Carcinoma ◽  
Chromosome 17 ◽  
Analysis Tool ◽  
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BACKGROUND Human epidermal growth factor receptor 2 (HER2) testing is routinely performed by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH) analyses for all new cases of invasive breast carcinoma. IHC is easier to perform, but analysis can be subjective and variable. FISH offers better diagnostic accuracy and added confidence, particularly when it is used to supplement weak IHC signals, but it is more labor intensive and costly than IHC. We examined the performance of droplet digital PCR (ddPCR) as a more precise and less subjective alternative for quantifying HER2 DNA amplification. METHODS Thirty-nine cases of invasive breast carcinoma containing ≥30% tumor were classified as positive or negative for HER2 by IHC, FISH, or both. DNA templates for these cases were prepared from formalin-fixed paraffin-embedded (FFPE) tissues to determine the HER2 copy number by ddPCR. ddPCR involved emulsifying hydrolysis probe–based PCR reaction mixtures containing the ERBB2 [v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian); also known as HER2] gene and chromosome 17 centromere assays into nanoliter-sized droplets for thermal cycling and analysis. RESULTS ddPCR distinguished, through differences in the level of HER2 amplification, the 10 HER2-positive samples from the 29 HER2-negative samples with 100% concordance to HER2 status obtained by FISH and IHC analysis. ddPCR results agreed with the FISH results for the 6 cases that were equivocal by IHC analyses, confirming 2 of these samples as positive for HER2 and the other 4 as negative. CONCLUSIONS ddPCR can be used as a molecular-analysis tool to precisely measure copy number alterations in FFPE samples of heterogeneous breast tumor tissue.

scholarly journals Detection of c-erbB-2 mRNAs Using Dig-Labelled Oligonucleotide Probe within situHybridisation in Human Breast Carcinoma: Comparison with Immunohistochemical Results

1998 ◽  
Vol 16 (4) ◽  
pp. 201-209 ◽  
Author(s):  
Melek Öztürk ◽  
Sema Bolkent ◽  
Selma Yilmazer ◽  
Gültekin Kaner ◽  
Hilal Ünal
Keyword(s):  
Gene Amplification ◽  
Prognostic Significance ◽  
Malignant Cell ◽  
Human Breast ◽  
Probe Concentration ◽  
Formalin Fixed Paraffin ◽  
Protein Overproduction ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Amplification and overexpression of the c-erbB-2 oncogene are of prognostic significance in human breast cancer. Overexpression of c-erbB-2 is the result of gene amplification. However, increased transcript levels of c-erbB-2 are also detected in the absence of gene amplification. In this study for the detection of the overexpression mRNAin situhybridisation (ISH) and immunohistochemistry (IHC) were used. Our aim was to develop the suitable mRNA ISH protocol for formalin-fixed paraffin-embedded material and to compare the localisation of transcripts and protein products in 20 primary breast carcinomas. Sections were immunostained with monoclonal c-erbB-2 antibody. In ISH method digoxigenin-labelled oligoprobe was used for the detection of c-erbB-2 mRNAs. We determined optimal condition for the ISH procedure (e.g., probe concentration, digestion, post washing). c-erbB-2 protein overproduction was detected in 11/20 cases with IHC. The mRNA signals were observed in malignant cell cytoplasm in 6/20 cases by ISH. ISH positive signals were found in only one case without detected overexpression of the protein. There were cell to cell variations in the hybridisation signals even within individual tumours. The ISH and IHC positive signals for c-erbB-2 was observed mostly in infiltrating ductal carcinomas that belong to aggressive lesions.

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