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Single cell analysis of cancer cells using an improved RT-MLPA method has potential for cancer diagnosis and monitoring
KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, Centres, Science for Life Laboratory, SciLifeLab.
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2015 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 5, 16519Article in journal (Refereed) Published
Abstract [en]

Single cell analysis techniques have great potential in the cancer genomics feld. The detection and characterization of circulating tumour cells are important for identifying metastatic disease at an early stage and monitoring it. This protocol is based on transcript profiling using Reverse Transcriptase Multiplex Ligation-dependent Probe Amplification (RT-MLPA), which is a specific method for simultaneous detection of multiple mRNA transcripts. Because of the small amount of (circulating) tumour cells, a pre-amplification reaction is performed after reverse transcription to generate a sufficient number of target molecules for the MLPA reaction. We designed a highly sensitive method for detecting and quantifying a panel of seven genes whose expression patterns are associated with breast cancer, and optimized the method for single cell analysis. For detection we used a fluorescence-dependent semi-quantitative method involving hybridization of unique barcodes to an array. We evaluated the method using three human breast cancer cell lines and identified specific gene expression profiles for each line. Furthermore, we applied the method to single cells and confirmed the heterogeneity of a cell population. Successful gene detection from cancer cells in human blood from metastatic breast cancer patients supports the use of RT-MLPA as a diagnostic tool for cancer genomics.

Place, publisher, year, edition, pages
Nature Publishing Group, 2015. Vol. 5, 16519
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:kth:diva-196159DOI: 10.1038/srep16519ISI: 000364461900003Scopus ID: 2-s2.0-84983542835OAI: oai:DiVA.org:kth-196159DiVA: diva2:1046980
Note

Funding Details: Knut and Alice Wallenberg Foundation. QC 20161116

Available from: 2016-11-16 Created: 2016-11-14 Last updated: 2017-02-28Bibliographically approved

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Kvastad, LindaWerne Solnestam, BeataJohansson, ErikSahlén, PelinVickovic, SanjaLundeberg, Joakim
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CiteExportLink to record
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Citation style
  • apa
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Output format
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