Open AccessIsolation and molecular analysis of circulating tumor cells from lung cancer patients using a microfluidic chip type cell sorter
Author(s) -
Watanabe Masaru,
Kenmotsu Hirotsugu,
Ko Ryo,
Wakuda Kazushige,
Ono Akira,
Imai Hisao,
Taira Tetsuhiko,
Naito Tateaki,
Murakami Haruyasu,
Abe Masato,
Endo Masahiro,
Nakajima Takashi,
Koh Yasuhiro,
Takahashi Toshiaki
Publication year - 2018
Publication title -
cancer science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 1347-9032
DOI - 10.1111/cas.13692
Subject(s) - lung cancer , circulating tumor cell , liquid biopsy , pathology , microfluidic chip , medicine , digital polymerase chain reaction , cancer research , cancer , biology , chip , polymerase chain reaction , gene , metastasis , computer science , telecommunications , biochemistry
Circulating tumor cells ( CTC s) are a tumor‐derived material utilized for liquid‐based biopsy; however, capturing rare CTC s for further molecular analysis remains technically challenging, especially in non‐small‐cell lung cancer. Here, we report the results of a clinical evaluation of On‐chip Sort, a disposable microfluidic chip‐based cell sorter, for capture and molecular analysis of CTC s from patients with lung adenocarcinoma. Peripheral blood was collected from 30 metastatic lung adenocarcinoma patients to enumerate CTC s using both On‐chip Sort and CellSearch in a blind manner. Captured cells by On‐chip Sort were subjected to further molecular analysis. Peripheral blood samples were also used for detection of EGFR mutations in plasma using droplet digital PCR . Significantly more CTC s were detected by On‐chip Sort (22/30; median 5; range, 0–18 cells/5 mL blood) than by CellSearch (9/30; median, 0; range, 0–12 cells/7.5 mL) ( P < 0.01). Thirteen of 30 patients who had a negative CTC count by CellSearch had a positive CTC count by On‐chip Sort. EGFR mutations in CTC s captured by On‐chip Sort were observed in 40.0% (8/20) of patients with EGFR ‐mutated primary tumor. EGFR mutations were often observed in 53.3% (8/15) of patients detected in plasma DNA . Expressions of EGFR and vimentin protein on CTC s were also successfully assessed using On‐chip Sort. These results suggest that On‐chip Sort is an efficient method to detect and capture rare CTC s from patients with lung adenocarcinoma that are undetectable with CellSearch. Mutation detection using isolated CTC s remains to be further tackled ( UMIN 000012488).