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Robustness of biomarker determination in breast cancer by RT-qPCR: impact of tumor cell content, DCIS and non-neoplastic breast tissue

التفاصيل البيبلوغرافية
العنوان: Robustness of biomarker determination in breast cancer by RT-qPCR: impact of tumor cell content, DCIS and non-neoplastic breast tissue
المؤلفون: Kornelia Schlombs, Hans-Anton Lehr, K Hartmann, Claudia Gürtler, Ugur Sahin, Marcus Schmidt, Mark Laible
المصدر: Diagnostic Pathology, Vol 13, Iss 1, Pp 1-10 (2018)
Diagnostic Pathology
بيانات النشر: Springer Science and Business Media LLC, 2018.
سنة النشر: 2018
مصطلحات موضوعية: 0301 basic medicine, Pathology, Tissue Fixation, Receptor, ErbB-2, Adipose tissue, Breast cancer, 0302 clinical medicine, skin and connective tissue diseases, Reverse Transcriptase Polymerase Chain Reaction, General Medicine, Prognosis, Real-time polymerase chain reaction, 030220 oncology & carcinogenesis, Tissue heterogeneity, Female, Receptors, Progesterone, lcsh:RB1-214, medicine.medical_specialty, Histology, DCIS, Non-neoplastic tissue, Breast Neoplasms, Laser Capture Microdissection, Biology, Real-Time Polymerase Chain Reaction, Pathology and Forensic Medicine, Fixatives, 03 medical and health sciences, Predictive Value of Tests, Formaldehyde, Biomarkers, Tumor, lcsh:Pathology, medicine, Carcinoma, Humans, RNA, Messenger, Research, Gene Expression Profiling, RT-qPCR, Estrogen Receptor alpha, Reproducibility of Results, Ductal carcinoma, medicine.disease, Fold change, Gene expression profiling, Carcinoma, Intraductal, Noninfiltrating, Ki-67 Antigen, MammaTyper, 030104 developmental biology, Tumor cell content, Macrodissection
الوصف: Background Tissue heterogeneity in formalin-fixed paraffin-embedded (FFPE) breast cancer specimens may affect the accuracy of reverse transcription quantitative real-time PCR (RT-qPCR). Herein, we tested the impact of tissue heterogeneity of breast cancer specimen on the RT-qPCR-based gene expression assay MammaTyper®. Methods MammaTyper® quantifies the mRNA expression of the four biomarkers ERBB2, ESR1, PGR, and MKI67. Based on pre-defined cut-off values, this molecular in vitro diagnostic assay permits binary marker classification and determination of breast cancer subtypes as defined by St Gallen 2013. In this study, we compared data from whole FFPE sections with data obtained in paired RNA samples after enrichment for invasive carcinoma via macro- or laser-capture micro-dissection. Results Compared to whole sections, removal of surrounding adipose tissue by macrodissection generated mean absolute 40-ddCq differences of 0.28–0.32 cycles for all four markers, with ≥90% concordant binary classifications. The mean raw marker Cq values in the adipose tissue were delayed by 6 to 7 cycles compared with the tumor-enriched sections, adding a trivial linear fold change of 1.0078 to 1.0156. Comparison of specimens enriched for invasive tumor with whole sections with as few as 20% tumor cell content resulted in mean absolute differences that remained on average below 0.59 Cq. The mean absolute difference between whole sections containing up to 60% ductal carcinoma in situ (DCIS) and specimens after dissection of DCIS was only 0.16–0.25 cycles, although there was a tendency for higher gene expression in DCIS. Observed variations were related to small size of samples and proximity of values to the limit of detection. Conclusion Expression of ESR1, PGR, ERBB2 and MKI67 by MammaTyper® is robust in clinical FFPE samples. Assay performance was unaffected by adipose tissue and was stable in samples with as few as 20% tumor cell content and up to 60% DCIS. Electronic supplementary material The online version of this article (10.1186/s13000-018-0760-6) contains supplementary material, which is available to authorized users.
تدمد: 1746-1596
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::821fd90a6d1887d5dc5fa720314c38c9
https://doi.org/10.1186/s13000-018-0760-6
حقوق: OPEN
رقم الأكسشن: edsair.doi.dedup.....821fd90a6d1887d5dc5fa720314c38c9
قاعدة البيانات: OpenAIRE
الوصف
تدمد:17461596