Abstract

Circulating tumour cells (CTCs) are important indicators of metastatic cancer and may provide critical information for individualized treatment. As CTCs are usually very rare, the techniques to obtain information from very small numbers of cells are crucial. Here, we propose a method to perform a single cell quantitative reverse transcription polymerase chain reaction (qPCR) analysis of rare tumour cells. We utilized a microfluidic immunomagnetic assay to separate cancer cells from blood. A combination of detailed immunofluorescence and laser microdissection enabled the precise selection of individual cells. Cancer cells that were spiked into blood were successfully separated and picked up for a single cell PCR analysis. The breast cancer cell lines MCF7, SKBR3 and MDAMB231 were tested with 10 different genes. The result of the single cell analysis matched the results from a few thousand cells. Some markers (e.g., ER, HER2) that are commonly used for cancer identification showed relatively large deviations in expression levels. However, others (e.g., GRB7) showed deviations that are small enough to supplement single cell disease profiling.

Details

Title
An Immunofluorescence-Assisted Microfluidic Single Cell Quantitative Reverse Transcription Polymerase Chain Reaction Analysis of Tumour Cells Separated from Blood
Author
Hoshino, Kazunori 1 ; Chung, HaeWon 2 ; Wu, Chun-Hsien 3 ; Rajendran, Kaarthik 3 ; Yu-Yen, Huang 4 ; Chen, Peng 3 ; Sokolov, Konstantin V 5 ; Kim, Jonghwan 2 ; Zhang, John XJ 4 

 Department of Biomedical Engineering, University of Connecticut, Storrs, CT, USA 
 Department of Molecular Biosciences, Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, TX, USA 
 Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, USA 
 Thayer School of Engineering, Dartmouth College, Hanover, NH, USA 
 Department of Imaging Physics, University of Texas MD Anderson Cancer Center, Houston, TX, USA 
Publication year
2015
Publication date
Jan 2015
Publisher
Sage Publications Ltd.
e-ISSN
18494544
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2179007532
Copyright
© 2015. This work is published under http://creativecommons.org/licenses/by/3.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.