MicroRNA for circulating tumor cells detection in breast cancer: In silico and in vitro analysis

e22027 Background: Detection of circulating tumor cells (CTC) may provide diagnostic and prognostic information in breast cancer (bc) patients. Deregulation of microRNAs (miRNAs) is frequent in tumors, including breast cancer. It is suggested miRNAs play a role in tumor progression. We hypothesized that miRNAs may constitute a promising new class of cancer biomarkers for CTC detection. Our objective has been to identify miRNAs potentially useful for CTC detection. Methods: Phase I preclinical study was performed by means of computational tools for miRNAs profiling including MIRGATOR, MIRBASE, SMIRNAdb, GeneHUB-GEPIS, MICRORNA.ORG, and MIRNAMAP. In silico data were used to identify and prioritize miRNAs highly expressed in breast cancer, but absent in hematopoietic-derived sources. Selected miRNAs were evaluated by means of qRT-PCR in breast cancer and Hematopoietic cell lines, normal blood, and blood from breast cancer patients. Results: Computational tools identify a set of miRNAs highly expressed in breast cancer sources in relation to hematopoietic samples. Among these were miR-141, miR-200c, miR-196a, miR-203, miR-200a, miR-200b, miR-32, miR-375, miR-31, miR-193a, and miR-205. For instance, relative expression of miR-32 was 104 higher in bc cell lines (N= 5) than in normal PB (N = 19) using qRT-PCR. In a preliminary analysis of PB from bc patients (stage IV) higher relative expression levels for selected miRNAs were found comparing with age-matched controls’ blood. Conclusions: Our results suggest that miRNA bioinformatic approach is a useful high-throughput method to select bc-associated miRNAS. The selected miRNAs should be further evaluated for their potential as markers for CTC detection. We next investigate if blood miRNA profile could predict tumor progression and survival. Supported by grants PI06/1541 and PI07/0477 from Fondo de Investigaciones Sanitarias (FIS), Instituto de Salud Carlos III. No significant financial relationships to disclose.

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Abstract P1-04-11: Recovery rates of circulating tumor cells in breast cancer cell lines spiked into peripheral blood

10.1158/0008-5472.sabcs13-p1-04-11 ◽

2013 ◽

Author(s):

W Zhu ◽

N Mineyev ◽

A Ring ◽

D Tripathy ◽

V Punj ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Cell Lines ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Peripheral Blood ◽

Cancer Cell Lines ◽

Breast Cancer Cell Lines ◽

Recovery Rates

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Circulating Tumor Cells: Detection Methods and Potential Clinical Application in Breast Cancer

Journal of Breast Cancer ◽

10.4048/jbc.2010.13.2.125 ◽

2010 ◽

Vol 13 (2) ◽

pp. 125 ◽

Cited By ~ 16

Author(s):

Seung Il Kim ◽

Hyo-il Jung

Keyword(s):

Breast Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Clinical Application ◽

Detection Methods ◽

Potential Clinical Application

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Circulating tumor cells (CTCs): Detection methods and their clinical relevance in breast cancer

Cancer Treatment Reviews ◽

10.1016/j.ctrv.2009.03.004 ◽

2009 ◽

Vol 35 (5) ◽

pp. 463-474 ◽

Cited By ~ 206

Author(s):

Bianca Mostert ◽

Stefan Sleijfer ◽

John A. Foekens ◽

Jan Willem Gratama

Keyword(s):

Breast Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Clinical Relevance ◽

Detection Methods

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Survivin gene expression in the primary tumor and circulating tumor cells – a new biomarker of tumor progression of breast cancer

Annals of Oncology ◽

10.1093/annonc/mdw363.20 ◽

2016 ◽

Vol 27 ◽

pp. vi21 ◽

Cited By ~ 1

Author(s):

Y. Shliakhtunou

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Tumor Progression ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Primary Tumor ◽

Survivin Gene

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Evolution of Estrogen Receptor Status from Primary Tumors to Metastasis and Serially Collected Circulating Tumor Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21082885 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2885 ◽

Cited By ~ 1

Author(s):

Carina Forsare ◽

Pär-Ola Bendahl ◽

Eric Moberg ◽

Charlotte Levin Tykjær Jørgensen ◽

Sara Jansson ◽

...

Keyword(s):

Breast Cancer ◽

Estrogen Receptor ◽

Tumor Progression ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Estrogen Receptor Status ◽

Metastatic Breast ◽

Primary Tumors ◽

Er Positive ◽

Er Status

Background: The estrogen receptor (ER) can change expression between primary tumor (PT) and distant metastasis (DM) in breast cancer. A tissue biopsy reflects a momentary state at one location, whereas circulating tumor cells (CTCs) reflect real-time tumor progression. We evaluated ER-status during tumor progression from PT to DM and CTCs, and related the ER-status of CTCs to prognosis. Methods: In a study of metastatic breast cancer, blood was collected at different timepoints. After CellSearch® enrichment, CTCs were captured on DropMount slides and evaluated for ER expression at baseline (BL) and after 1 and 3 months of therapy. Comparison of the ER-status of PT, DM, and CTCs at different timepoints was performed using the McNemar test. The primary endpoint was progression-free survival (PFS). Results: Evidence of a shift from ER positivity to negativity between PT and DM was demonstrated (p = 0.019). We found strong evidence of similar shifts from PT to CTCs at different timepoints (p < 0.0001). ER-positive CTCs at 1 and 3 months were related to better prognosis. Conclusions: A shift in ER-status from PT to DM/CTCs was demonstrated. ER-positive CTCs during systemic therapy might reflect the retention of a favorable phenotype that still responds to therapy.

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Detection Methods and Clinical Applications of Circulating Tumor Cells in Breast Cancer

Frontiers in Oncology ◽

10.3389/fonc.2021.652253 ◽

2021 ◽

Vol 11 ◽

Author(s):

Hongyi Zhang ◽

Xiaoyan Lin ◽

Yuan Huang ◽

Minghong Wang ◽

Chunmei Cen ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Cancer Survival ◽

Survival Rates ◽

Circulatory System ◽

Clinical Applications ◽

Detection Methods ◽

Sample Collection ◽

Detection And Diagnosis

Circulating Tumor Cells (CTCs) are cancer cells that split away from the primary tumor and appear in the circulatory system as singular units or clusters, which was first reported by Dr. Thomas Ashworth in 1869. CTCs migrate and implantation occurs at a new site, in a process commonly known as tumor metastasis. In the case of breast cancer, the tumor cells often migrate into locations such as the lungs, brain, and bones, even during the early stages, and this is a notable characteristic of breast cancer. Survival rates have increased significantly over the past few decades because of progress made in radiology and tissue biopsy, making early detection and diagnosis of breast cancer possible. However, liquid biopsy, particularly that involving the collection of CTCs, is a non-invasive method to detect tumor cells in the circulatory system, which can be easily isolated from human plasma, serum, and other body fluids. Compared to traditional tissue biopsies, fluid sample collection has the advantages of being readily available and more acceptable to the patient. It can also detect tumor cells in blood earlier and in smaller numbers, possibly allowing for diagnosis prior to any tumor detection using imaging methods. Because of the scarcity of CTCs circulating in blood vessels (only a few CTCs among billions of erythrocytes and leukocytes), thorough but accurate detection methods are particularly important for further clinical applications.

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Gene Expression Profiling of Circulating Tumor Cells in Breast Cancer

Clinical Chemistry ◽

10.1373/clinchem.2014.229476 ◽

2015 ◽

Vol 61 (1) ◽

pp. 278-289 ◽

Cited By ~ 15

Author(s):

Emanuela Fina ◽

Maurizio Callari ◽

Carolina Reduzzi ◽

Francesca D'Aiuto ◽

Gabriella Mariani ◽

...

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Cell Lines ◽

Magnetic Beads ◽

Expression Profiles ◽

Clinical Samples ◽

Breast Cancer Cell Lines ◽

Tissue Samples

Abstract BACKGROUND Determining the transcriptional profile of circulating tumor cells (CTCs) may allow the acquisition of clinically relevant information while overcoming tumor heterogeneity-related biases associated with use of tissue samples for biomarker assessment. However, such molecular characterization is challenging because CTCs are rare and outnumbered by blood cells. METHODS Here, we describe a technical protocol to measure the expression of >29 000 genes in CTCs captured from whole blood with magnetic beads linked with antibodies against epithelial cell adhesion molecule (EpCAM) and the carcinoma-associated mucin, MUC1, designed to be used for CTC characterization in clinical samples. Low numbers of cells (5–200) from the MCF7 and MDA-MB-468 breast cancer cell lines were spiked in healthy donor blood samples and isolated with the AdnaTest EMT-1/Stem CellSelect kit. Gene expression profiles (GEPs) were obtained with the WG-DASL HT assay and compared with GEPs obtained from RNA isolated from cultured cell lines and unspiked samples. RESULTS GEPs from samples containing 25 or more spiked cells correlated (r = 0.95) with cognate 100-ng RNA input samples, clustered separately from blood control samples, and allowed MCF7 and MDA-MB-468 cells to be distinguished. GEPs with comparable technical quality were also obtained in a preliminary series of clinical samples. CONCLUSIONS Our approach allows technically reliable GEPs to be obtained from isolated CTCs for the acquisition of biologically useful information. It is reproducible and suitable for application in prospective studies to assess the clinical utility of CTC GEPs, provided that >25 CTCs can be isolated.

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