Subtyping of Circulating Tumor Cells Using a Sequential Staining Method
Date Issued
2015
Date
2015
Author(s)
Chang, Po-Kang
Abstract
According to Globocan, there were 8.2 million cancer deaths in 2012 worldwide, and metastasis was considered to be related to the death of many cancer patients. Nowadays circulating tumor cells (CTCs) obtained from blood samples has been considered as a promising indicator for prognosis such as progression-free survival (PFS) and overall survival (OS) of patients, and has been used and validated in many clinical trials in metastatic breast, prostate and colorectal cancers. Therefore, many technologies have been developed for CTCs enrichment and isolation. However, several studies show that CTC enumeration alone is not enough for clinical utility due to their high heterogeneity. Therefore, further analysis of enumerated CTCs has become increasingly important. This work presents a semi-automated sequential staining system combined with a single cell retrieval system as a competent multi-functional platform. It redesigned the staining procedure in order to obtain clearer signals and get more information about the phenotype of CTCs. Since in most cases, laboratories have only four-channel-fluorescence microscopes, where PE signal is likely to interfere with FITC signal due to its light leakage problem. Moreover, three channels may be used to identify CTCs, and only one channel remains for further information. By using this system, we may obtain five fluorescent signals in a four-channel-fluorescence microscope for both CTC detection and its subtype identification. Experiments characterized MCF-7, AU565, and MDA-MB-231 and used total about 50 cells to simulate ER/HER2 subtypes with different ratio of cell lines. Results show that recovery rate and staining efficiency in sequential staining system has no significant differences to the disk system. Also, it indicates that the performances were good for HER2 expression, while about SD=12.0% for ER expression. The overall proportion of mixed subtypes were successfully predicted and detected. With this multi-functional platform, we were able to provide more information about the subtype expression and may offer further analyses on CTCs.
Subjects
Circulating tumor cells
subtype
sequential staining
SDGs
Type
thesis
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