期刊
TALANTA
卷 185, 期 -, 页码 229-236出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2018.03.083
关键词
Circulating tumor cells (CTCs); Immuno-magnetic beads; Epithelial protein; Lung cancer; EGFR mutant; Droplet digital PCR (ddPCR)
资金
- National Natural Science Foundation of China [81472751, 61571428, 61571429]
- Program of Science and Technology Commission of Shanghai Municipality [17JC1401001]
- Key Research Program of China [2017YFA0205303]
Circulating tumor cells (CTCs) have become an important biomarker for liquid biopsy to monitor tumor progression and indicate response to therapies. Many epithelial cellular adhesion molecule (EpCAM) dependent CTC isolation methods have been developed, which have a limitation for low EpCAM expressed tumor cells. In an effort to overcome these drawbacks, we developed combined immunomagnetic beads (EpCAM, Mucin1 and epidermal growth factor receptor) to sensitively isolate CTCs for immunofluorescence analysis and genetic characterization. With this combined immunomagnetic beads, 93.35% H446 cells from spiked blood sample can be recovered. We were able to detect CTCs in 127 among 143 patients included in the study (88.8%). Some CTC clusters were captured with the combined magnetic beads system. In 17 of them, CTCs after chemotherapy significantly decreased compared to that before chemotherapy (4.42 (+/- 3.94) vs. 12 (+/- 7)/mL, P = 0.002). For subsequent genetic characterization of CTCs, 2 of 6 samples, using a droplet digital PCR (ddPCR) chip, have detectable EGFR L858R mutation in the cells enriched with the combined immunomagnetic beads. In conclusion, this method integrating the combined immunomagnetic beads and the ddPCR chip for CTCs detection can be of potential application in terms of diagnosis, therapeutic evaluation and personalized medicine in lung cancer.
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