title: Classification of circulating tumor cells by epithelial-mesenchymal transition markers.
abstract: In cancer, epithelial-mesenchymal transition (EMT) is associated with metastasis. Characterizing EMT phenotypes in circulating tumor cells (CTCs) has been challenging because epithelial marker-based methods have typically been used for the isolation and detection of CTCs from blood samples. The aim of this study was to use the optimized CanPatrol CTC enrichment technique to classify CTCs using EMT markers in different types of cancers. The first step of this technique was to isolate CTCs via a filter-based method; then, an RNA in situ hybridization (RNA-ISH) method based on the branched DNA signal amplification technology was used to classify the CTCs according to EMT markers. Our results indicated that the efficiency of tumor cell recovery with this technique was at least 80%. When compared with the non-optimized method, the new method was more sensitive and more CTCs were detected in the 5-ml blood samples. To further validate the new method, 164 blood samples from patients with liver, nasopharyngeal, breast, colon, gastric cancer, or non-small-cell lung cancer (NSCLC) were collected for CTC isolation and characterization. CTCs were detected in 107 (65%) of 164 blood samples, and three CTC subpopulations were identified using EMT markers, including epithelial CTCs, biophenotypic epithelial/mesenchymal CTCs, and mesenchymal CTCs. Compared with the earlier stages of cancer, mesenchymal CTCs were more commonly found in patients in the metastatic stages of the disease in different types of cancers. Circulating tumor microemboli (CTM) with a mesenchymal phenotype were also detected in the metastatic stages of cancer. Classifying CTCs by EMT markers helps to identify the more aggressive CTC subpopulation and provides useful evidence for determining an appropriate clinical approach. This method is suitable for a broad range of carcinomas.
authors.affiliation: SurExam Bio-Tech, Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China.
authors.forename: Shiyang
authors.initials: S
authors.lastname: Wu
authors.affiliation: SurExam Bio-Tech, Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China.
authors.forename: Suyan
authors.initials: S
authors.lastname: Liu
authors.affiliation: SurExam Bio-Tech, Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Liu
authors.affiliation: SurExam Bio-Tech, Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China.
authors.forename: Jiefeng
authors.initials: J
authors.lastname: Huang
authors.affiliation: SurExam Bio-Tech, Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Pu
authors.affiliation: Oncology Department, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou, P.R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Hepatobiliary Surgery, Guangzhou Nanfang Hospital, Guangzhou, P.R. China.
authors.forename: Dinghua
authors.initials: D
authors.lastname: Yang
authors.affiliation: Department of General Surgery, Guangzhou Nanfang Hospital, Guangzhou, P.R. China.
authors.forename: Haijun
authors.initials: H
authors.lastname: Deng
authors.affiliation: Department of Pathology, The First Hospital Affiliated with Guangdong College of Pharmacy, Guangzhou, P.R. China.
authors.forename: Ning
authors.initials: N
authors.lastname: Yang
authors.affiliation: SurExam Bio-Tech, Guangzhou Technology Innovation Base, 80 Lan Yue Road, Science City, Guangzhou, P.R. China.
authors.forename: Jiasen
authors.initials: J
authors.lastname: Xu
mesh_terms: D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D005260:Female; D056945:Hep G2 Cells; D006801:Humans; D017403:In Situ Hybridization; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D009369:Neoplasms / Q000175:diagnosis* / Q000235:genetics; D009360:Neoplastic Cells, Circulating / Q000378:metabolism* / Q000473:pathology*; D015203:Reproducibility of Results
chemical_list: D014408:Biomarkers, Tumor