title: Sensitive cytometry based system for enumeration, capture and analysis of gene mutations of circulating tumor cells.
abstract: Methods for the enumeration and molecular characterization of circulating tumor cells (CTC) have been actively investigated. However, such methods are still technically challenging. We have developed a novel epithelial cell adhesion molecule independent CTC enumeration system integrated with a sorting system using a microfluidics chip. We compared the number of CTC detected using our system with those detected using the CellSearch system in 46 patients with various cancers. We also evaluated epidermal growth factor receptor (EGFR) and PIK3CA mutations of captured CTC in a study of 4 lung cancer and 4 breast cancer patients. The percentage of samples with detected CTC was significantly higher with our system (65.2%) than with CellSearch (28.3%). The number of detected CTC per patient using our system was statistically higher than that using CellSearch (median 5, 0; P = 0.000172, Wilcoxon test). In the mutation analysis study, the number of detected CTC per patient was low (median for lung, 4.5; median for breast, 5.5); however, it was easy to detect EGFR and PIK3CA mutations in the CTC of 2 lung and 1 breast cancer patient, respectively, using a commercially available kit. Our system is more sensitive than CellSearch in CTC enumeration of various cancers and is also capable of detecting EGFR and PIK3CA mutations in the CTC of lung and breast cancer patients, respectively.
authors.affiliation: Shien-Lab, National Cancer Center Hospital, Tokyo, Japan.|Medical Oncology/Hematology, Kobe University Graduate School of Medicine, Kobe, Japan.|Department of Medical Oncology, Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital, Tokyo, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sawada
authors.affiliation: Drug Discovery and Development Division, Shizuoka Cancer Center Research Institute, Nagaizumi, Japan.|Third Department of Internal Medicine, Wakayama Medical University, Wakayama, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Watanabe
authors.affiliation: On-Chip Biotechnologies, Tokyo, Japan.
authors.forename: Yuu
authors.initials: Y
authors.lastname: Fujimura
authors.affiliation: Shien-Lab, National Cancer Center Hospital, Tokyo, Japan.
authors.forename: Shigehiro
authors.initials: S
authors.lastname: Yagishita
authors.affiliation: Department of Medical Oncology, Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital, Tokyo, Japan.
authors.forename: Tatsu
authors.initials: T
authors.lastname: Shimoyama
authors.affiliation: Department of Medical Oncology, Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital, Tokyo, Japan.
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Maeda
authors.affiliation: Division of Internal Medicine and Thoracic Oncology, National Cancer Center Hospital, Tokyo, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Kanda
authors.affiliation: Division of Breast and Medical Oncology, National Cancer Center Hospital, Tokyo, Japan.
authors.forename: Mayu
authors.initials: M
authors.lastname: Yunokawa
authors.affiliation: Division of Breast and Medical Oncology, National Cancer Center Hospital, Tokyo, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Tamura
authors.affiliation: Division of Internal Medicine and Thoracic Oncology, National Cancer Center Hospital, Tokyo, Japan.
authors.forename: Tomohide
authors.initials: T
authors.lastname: Tamura
authors.affiliation: Medical Oncology/Hematology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Hironobu
authors.initials: H
authors.lastname: Minami
authors.affiliation: Drug Discovery and Development Division, Shizuoka Cancer Center Research Institute, Nagaizumi, Japan.|Third Department of Internal Medicine, Wakayama Medical University, Wakayama, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Koh
authors.affiliation: Shien-Lab, National Cancer Center Hospital, Tokyo, Japan.|Department of Laboratory Medicine, Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital, Tokyo, Japan.
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Koizumi
mesh_terms: D000328:Adult; D000368:Aged; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000235:genetics / Q000556:secondary; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000556:secondary; D016022:Case-Control Studies; D002452:Cell Count; D045744:Cell Line, Tumor; D002469:Cell Separation; D058534:Class I Phosphatidylinositol 3-Kinases; D004252:DNA Mutational Analysis; D066246:ErbB Receptors / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D056656:Lab-On-A-Chip Devices; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics; D017384:Sequence Deletion
chemical_list: D019869:Phosphatidylinositol 3-Kinases; D058534:Class I Phosphatidylinositol 3-Kinases; C484760:PIK3CA protein, human; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: CTC; Cell sorter; EGFR; Flow cytometry; PIK3CA; Scorpion ARMS