Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot Study

<i>Background and Objectives:</i> Breast cancer (BC) remains one of the major causes of cancer death in women worldwide. The difficulties in assessing the deep molecular mechanisms involved in this pathology arise from its high complexity and diverse tissue subtypes. Long non-coding RNAs...

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Main Authors: Anca Marcu, Diana Nitusca, Adrian Vaduva, Flavia Baderca, Natalia Cireap, Dorina Coricovac, Cristina Adriana Dehelean, Edward Seclaman, Razvan Ilina, Catalin Marian
Format: Article
Language:English
Published: MDPI AG 2021-04-01
Series:Medicina
Subjects:
Online Access:https://www.mdpi.com/1648-9144/57/4/371
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spelling doaj-ffca53d67f504b75ab0de2e949225a5c2021-04-12T23:01:28ZengMDPI AGMedicina1010-660X1648-91442021-04-015737137110.3390/medicina57040371Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot StudyAnca Marcu0Diana Nitusca1Adrian Vaduva2Flavia Baderca3Natalia Cireap4Dorina Coricovac5Cristina Adriana Dehelean6Edward Seclaman7Razvan Ilina8Catalin Marian9Department of Biochemistry and Pharmacology, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr.2, 300041 Timişoara, RomaniaDepartment of Biochemistry and Pharmacology, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr.2, 300041 Timişoara, RomaniaDepartment of Microscopic Morphology, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr.2, 300041 Timişoara, RomaniaDepartment of Microscopic Morphology, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr.2, 300041 Timişoara, RomaniaDepartment of Surgical Oncology, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr.2, 300041 Timişoara, RomaniaFaculty of Pharmacy, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr. 2, 300041 Timişoara, RomaniaFaculty of Pharmacy, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr. 2, 300041 Timişoara, RomaniaDepartment of Biochemistry and Pharmacology, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr.2, 300041 Timişoara, RomaniaDepartment of Surgical Oncology, Municipal Hospital, Str. Gheorghe Dima Nr.5, 300254 Timişoara, RomaniaDepartment of Biochemistry and Pharmacology, Victor Babeş University of Medicine and Pharmacy, Pta Eftimie Murgu Nr.2, 300041 Timişoara, Romania<i>Background and Objectives:</i> Breast cancer (BC) remains one of the major causes of cancer death in women worldwide. The difficulties in assessing the deep molecular mechanisms involved in this pathology arise from its high complexity and diverse tissue subtypes. Long non-coding RNAs (lncRNAs) were shown to have great tissue specificity, being differentially expressed within the BC tissue subtypes. <i>Materials and Methods:</i> Herein, we performed lncRNA profiling by PCR array in triple negative breast cancer (TNBC) and luminal A tissue samples from 18 BC patients (nine TNBC and nine luminal A), followed by individual validation in BC tissue and cell lines. Tissue samples were previously archived in formalin-fixed paraffin-embedded (FFPE) samples, and the areas of interest were dissected using laser capture microdissection (LCM) technology. <i>Results:</i> Two lncRNAs (OTX2-AS1 and SOX2OT) were differentially expressed in the profiling analysis (fold change of 205.22 and 0.02, respectively, <i>p</i> < 0.05 in both cases); however, they did not reach statistical significance in the individual validation measurement (<i>p</i> > 0.05) when analyzed with specific individual assays. In addition, GAS5 and NEAT1 lncRNAs were individually assessed as they were previously described in the literature as being associated with BC. GAS5 was significantly downregulated in both TNBC tissues and cell lines compared to luminal A samples, while NEAT1 was significantly downregulated only in TNBC cells vs. luminal A. <i>Conclusions:</i> Therefore, we identified GAS5 lncRNA as having a differential expression in TNBC tissues and cells compared to luminal A, with possible implications in the molecular mechanisms of the TNBC subtype. This proof of principle study also suggests that LCM could be a useful technique for limiting the sample heterogeneity for lncRNA gene expression analysis in BC FFPE tissues. Future studies of larger cohort sizes are needed in order to assess the biomarker potential of lncRNA GAS5 in BC.https://www.mdpi.com/1648-9144/57/4/371breast cancertissue specificitylncRNAlaser capture microdissection
collection DOAJ
language English
format Article
sources DOAJ
author Anca Marcu
Diana Nitusca
Adrian Vaduva
Flavia Baderca
Natalia Cireap
Dorina Coricovac
Cristina Adriana Dehelean
Edward Seclaman
Razvan Ilina
Catalin Marian
spellingShingle Anca Marcu
Diana Nitusca
Adrian Vaduva
Flavia Baderca
Natalia Cireap
Dorina Coricovac
Cristina Adriana Dehelean
Edward Seclaman
Razvan Ilina
Catalin Marian
Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot Study
Medicina
breast cancer
tissue specificity
lncRNA
laser capture microdissection
author_facet Anca Marcu
Diana Nitusca
Adrian Vaduva
Flavia Baderca
Natalia Cireap
Dorina Coricovac
Cristina Adriana Dehelean
Edward Seclaman
Razvan Ilina
Catalin Marian
author_sort Anca Marcu
title Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot Study
title_short Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot Study
title_full Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot Study
title_fullStr Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot Study
title_full_unstemmed Long Non-Coding RNA Expression in Laser Micro-Dissected Luminal A and Triple Negative Breast Cancer Tissue Samples—A Pilot Study
title_sort long non-coding rna expression in laser micro-dissected luminal a and triple negative breast cancer tissue samples—a pilot study
publisher MDPI AG
series Medicina
issn 1010-660X
1648-9144
publishDate 2021-04-01
description <i>Background and Objectives:</i> Breast cancer (BC) remains one of the major causes of cancer death in women worldwide. The difficulties in assessing the deep molecular mechanisms involved in this pathology arise from its high complexity and diverse tissue subtypes. Long non-coding RNAs (lncRNAs) were shown to have great tissue specificity, being differentially expressed within the BC tissue subtypes. <i>Materials and Methods:</i> Herein, we performed lncRNA profiling by PCR array in triple negative breast cancer (TNBC) and luminal A tissue samples from 18 BC patients (nine TNBC and nine luminal A), followed by individual validation in BC tissue and cell lines. Tissue samples were previously archived in formalin-fixed paraffin-embedded (FFPE) samples, and the areas of interest were dissected using laser capture microdissection (LCM) technology. <i>Results:</i> Two lncRNAs (OTX2-AS1 and SOX2OT) were differentially expressed in the profiling analysis (fold change of 205.22 and 0.02, respectively, <i>p</i> < 0.05 in both cases); however, they did not reach statistical significance in the individual validation measurement (<i>p</i> > 0.05) when analyzed with specific individual assays. In addition, GAS5 and NEAT1 lncRNAs were individually assessed as they were previously described in the literature as being associated with BC. GAS5 was significantly downregulated in both TNBC tissues and cell lines compared to luminal A samples, while NEAT1 was significantly downregulated only in TNBC cells vs. luminal A. <i>Conclusions:</i> Therefore, we identified GAS5 lncRNA as having a differential expression in TNBC tissues and cells compared to luminal A, with possible implications in the molecular mechanisms of the TNBC subtype. This proof of principle study also suggests that LCM could be a useful technique for limiting the sample heterogeneity for lncRNA gene expression analysis in BC FFPE tissues. Future studies of larger cohort sizes are needed in order to assess the biomarker potential of lncRNA GAS5 in BC.
topic breast cancer
tissue specificity
lncRNA
laser capture microdissection
url https://www.mdpi.com/1648-9144/57/4/371
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