scholarly journals Comparing MicroRNA Profilings of Purified HER-2-Negative and HER-2-Positive Cells Validates miR-362-5p/Sema3A as Characteristic Molecular Change in Triple-Negative Breast Cancers

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Xiaoqing Zhang ◽  
Qi He ◽  
Leiqin Sun ◽  
Yanfei Zhang ◽  
Shengying Qin ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Protein Expression ◽  
Real Time ◽  
Differential Expression ◽  
Real Time Pcr ◽  
Therapeutic Target ◽  
Triple Negative ◽  
Quantitative Real Time Pcr ◽  
Her 2 ◽  
Tnbc Subtype

Background. HER-2 is a key molecule serving as the therapeutic target, prognostic biomarker, and classification marker in breast cancer. Accurate microRNA profilings had not been conducted in purified tumor cells of HER-2-negative and HER-2-positive tissue specimens obtained from breast cancer patients. Methods. (i) Differential expression microRNA discovery using laser capture microdissection- (LCM-) assisted specimen preparation and microRNA array chips on HER-2 overexpressing and triple-negative breast carcinoma (TNBC) subtype tissues, (ii) differential expression microRNA validation by quantitative real-time PCR, and (iii) independent validation on tissue microarray. Results. Five microRNAs (miR-20a-5p, miR-221-3p, miR-362-5p, miR-502-3p, and miR-222-3p) were screened and validated as upregulated microRNAs in TNBC cells comparing to HER-2 overexpressing cells using a microRNA array (5 cases in each group) and quantitative real-time PCR (20 cases in each group). The expression difference of miR-362-5p had the most significant statistical significance (p=0.0016) among the five microRNAs. The expression of miR-362-5p and its target gene Sema3A was further analyzed using in situ hybridization (ISH) and immunohistochemistry on standard tissue sections (n=150). 70.8% of HER-2-negative cells showed moderate expression of miR-362-5p whereas 20.4% HER-2-negative cells correlated with strong expression of miR-362-5p (p<0.0001). The proportion of patients with moderate/strong miR-362-5p expression in luminal, HER-2 overexpressing, and TNBC subtypes were 53.2%, 22.2%, and 74.3%, respectively (p=0.0002). High miR-362-5p expressers had shorter overall survival in the univariate analysis (p=0.046). There was a significant negative correlation between miR-362-5p and Sema3A expression (p<0.0001). The patients with negative/weak Sema3A protein expression had poorer prognosis than those with moderate (HR: 3.723, p=0.021) or strong (HR: 3.966, p=0.013) Sema3A protein expression in the multivariate analysis. Conclusions. miR-362-5p/Sema3A might provide a promising therapeutic pathway and represents a candidate therapeutic target of the TNBC subtype.

scholarly journals HER-2/neu Gene Copy Number Quantified by Real-Time PCR: Comparison of Gene Amplification, Heterozygosity, and Immunohistochemical Status in Breast Cancer Tissue

2003 ◽  
Vol 49 (2) ◽  
pp. 219-229 ◽  
Author(s):  
Melanie Königshoff ◽  
Jochen Wilhelm ◽  
Rainer M Bohle ◽  
Alfred Pingoud ◽  
Meinhard Hahn
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Cell Lines ◽  
Real Time Pcr ◽  
Gene Copy ◽  
Protein Overexpression ◽  
Tissue Samples ◽  
Cancer Pathogenesis ◽  
Her 2 ◽  
Gene Status

Abstract Background: Amplification of the oncogene HER-2/neu influences breast cancer pathogenesis, and therapy and prognosis may be affected by the degree of amplification. The extent of amplification or protein overexpression typically is analyzed by fluorescence in situ hybridization or immunohistochemistry (IHC), but quantitative PCR techniques have been described that may provide alternatives to these methods. Methods: We developed a rapid-cycle, real-time PCR assay for quantification of HER-2/neu gene status. We compared results obtained with this assay with short tandem repeat findings by capillary electrophoresis (CE) and with protein overexpression assessments by IHC. Accuracy and linearity were tested on cell lines and with simulation experiments. We analyzed the amplification of HER-2/neu in 51 clinical tissue samples from patients with suspected breast cancer. Results: The intra- and interrun CVs for HER-2/neu quantification by real-time PCR were 12% and 18%, and the CV for different simulated amplification and deletion experiments was &lt;7%. The results for HER-2/neu gene status in cell lines matched the values reported in literature. We detected HER-2/neu amplification by real-time PCR in 11 samples, all from patients with invasive ductal carcinoma. Allelic imbalances were found by CE analyses in three samples and by protein overexpression in six samples; five of these were also detected by real-time PCR. Comparison of the quantification results with known prognostic indices yielded results similar to those reported in several other published studies. Conclusions: The assay is suitable for accurate and precise quantification of HER-2/neu copy numbers in tumor tissue samples obtained in routine clinical practice.

Preoperative assessment of HER-2/neu status in breast carcinoma: The role of quantitative real-time PCR on core-biopsy specimens

2010 ◽  
Vol 116 (2) ◽  
pp. 234-239 ◽  
Author(s):  
Tommaso Susini ◽  
Cecilia Bussani ◽  
Giulia Marini ◽  
Jacopo Nori ◽  
Simone Olivieri ◽  
...  
Keyword(s):  
Breast Carcinoma ◽  
Real Time ◽  
Real Time Pcr ◽  
Core Biopsy ◽  
Preoperative Assessment ◽  
Quantitative Real Time Pcr ◽  
Biopsy Specimens ◽  
Her 2

Borderline HER‐2 breast cancer cases: Histochemical versus real‐time PCR analysis and impact of different cut‐off values

2007 ◽  
Vol 67 (4) ◽  
pp. 402-412 ◽  
Author(s):  
G. Monego ◽  
V. Arena ◽  
N. Maggiano ◽  
L. Costarelli ◽  
A. Crescenzi ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Real Time Pcr ◽  
Pcr Analysis ◽  
Her 2

Evaluation of cisplatin on CD82 gene in breast cancer MCF-7 cells using quantitative real-time PCR

2011 ◽  
Vol 44 (13) ◽  
pp. S203
Author(s):  
Mokhtari Mohammad Javad ◽  
Hosseinian Zhilla ◽  
Akbarzadeh Azim ◽  
Kamyab Ahmad Reza ◽  
Ghasemi Soheil ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Mcf 7

Current Diagnostic Methods of HER-2/neu Detection in Breast Cancer With Special Regard to Real-Time PCR

2003 ◽  
Vol 27 (12) ◽  
pp. 1565-1570 ◽  
Author(s):  
Sabine Merkelbach-Bruse ◽  
Eva Wardelmann ◽  
Peter Behrens ◽  
Inge Losen ◽  
Reinhard Buettner ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Real Time Pcr ◽  
Diagnostic Methods ◽  
Special Regard ◽  
Her 2

PTEN expression in Palestinian women with breast cancer, particularly in triple-negative subtype.

2011 ◽  
Vol 29 (27_suppl) ◽  
pp. 27-27
Author(s):  
A. A. Khatib ◽  
M. N. AL Abed
Keyword(s):  
Breast Cancer ◽  
Protein Expression ◽  
Triple Negative ◽  
Pten Expression ◽  
High Grade ◽  
Advanced Stage ◽  
Pten Protein ◽  
Pten Loss ◽  
Palestinian Women ◽  
Her 2

27 Background: According to data from the Palestinian Ministry of Health, breast cancer is the most common malignancy affecting Palestinian women, being 30% of all cancers affecting female patients, 60% of them present with metastasis, very few studies were conducted to explore characteristics of this cancer in this population. Triple-negative breast cancer (TNBC) has distinct clinical and pathologic features. The fact that this type is more aggressive despite lacking the expression of growth promotion receptors, suggest other players, as defect in tumor suppressors such as PTEN gene. To our knowledge this is the first study in English literature correlating TNBC to loss of PTEN protein expression. The aim of this study is to establish data about the clinicopathological characteristics of breast cancer in Palestinian women, and to study the status of PTEN protein expression by immunohistochemistry in breast cancer and specifically in TNBC. Methods: 100 confirmed cases of breast cancer, were collected from different pathology centers, clinicopathological data; age, grade, stage for each case were specified. Each case was evaluated for the expression of hormone receptors, Her-2 status, and for PTEN protein expression by immunohistochemistry (IHC) to see if a certain pattern is noted. Results: Out of the 100 cases, the mean age at presentation was 53.3 years, 69.84% of the cases presented with advanced stage and 53% were high grade, 58% were ER positive and 46% were PR positive. HER-2 positive breast cancer were 26%. 30% of all breast cancer cases were TNBC subtype. Loss of PTEN expression was seen in 44% of cases. 60% of TNBC cases lost PTEN expression, which was statistically significant (p + 0.035), no significant correlation between PTEN loss and ER, PR, HER-2, grade or stage. Conclusions: Palestinian women have high incidence of TNBC in comparison to White population, and presented with high grade, and advanced stage tumors. There is a significant correlation between PTEN loss and TNBC that needs more research and study.

scholarly journals Simultaneous Quantitative Detection of Relevant Biomarkers in Breast Cancer by Quantitative Real-Time PCR

2006 ◽  
Vol 21 (1) ◽  
pp. 30-39 ◽  
Author(s):  
M. Labuhn ◽  
V. Vuaroqueaux ◽  
F. Fina ◽  
A. Schaller ◽  
I. Nanni-Metellus ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Mrna Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Detection ◽  
Mrna Levels ◽  
Quantitative Real Time Pcr ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Mrna Expression Levels

The assessment of ERα, PgR and HER2 status is routinely performed today to determine the endocrine responsiveness of breast cancer samples. Such determination is usually accomplished by means of immunohistochemistry and in case of HER2 amplification by means of fluorescent in situ hybridization (FISH). The analysis of these markers can be improved by simultaneous measurements using quantitative real-time PCR (Qrt-PCR). In this study we compared Qrt-PCR results for the assessment of mRNA levels of ERα, PgR, and the members of the human epidermal growth factor receptor family, HER1, HER2, HER3 and HER4. The results were obtained in two independent laboratories using two different methods, SYBR Green I and TaqMan probes, and different primers. By linear regression we demonstrated a good concordance for all six markers. The quantitative mRNA expression levels of ERα, PgR and HER2 also strongly correlated with the respective quantitative protein expression levels prospectively detected by EIA in both laboratories. In addition, HER2 mRNA expression levels correlated well with gene amplification detected by FISH in the same biopsies. Our results indicate that both Qrt-PCR methods were robust and sensitive tools for routine diagnostics and consistent with standard methodologies. The developed simultaneous assessment of several biomarkers is fast and labor effective and allows optimization of the clinical decision-making process in breast cancer tissue and/or core biopsies.

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