Detection of Circulating Tumor Cells and Circulating Tumor Microemboli in Gastric Cancer Xiumei Zheng, Li Fan, Pengfei Zhou, Hong Ma, Shaoyi Huang, Dandan.

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Detection of Circulating Tumor Cells and Circulating Tumor Microemboli in Gastric Cancer Xiumei Zheng, Li Fan, Pengfei Zhou, Hong Ma, Shaoyi Huang, Dandan Yu, Lei Zhao, Shengli Yang, Jun Liu, Ai Huang, Congli Cai, Xiaomeng Dai, Tao Zhang Translational Oncology Volume 10, Issue 3, Pages 431-441 (June 2017) DOI: 10.1016/j.tranon.2017.02.007 Copyright © 2017 The Authors Terms and Conditions

Figure 1 Cells enriched by the ISET method in patients with gastric cancer. Diff-Quick staining. (A) CTC: irregular nucleus; nuclear diameter >15μm; nuclear-cytoplasmic ratio >0.8; hyperchromatic nucleus and nonhomogeneous staining. (B) CTM: presence of tumor cell aggregation, prominent nucleoli seen in the nuclei. (C) Nucleus without plasma. (D) Normal blood cells isolated on the membrane. The scale bar represents 20μm. Translational Oncology 2017 10, 431-441DOI: (10.1016/j.tranon.2017.02.007) Copyright © 2017 The Authors Terms and Conditions

Figure 2 Immunofluorescent staining of CTCs/CTM isolated by ISET assay. (A–D) Stained with CK8/18/19 (green fluorescence for epithelial cells), Vimentin (purple fluorescence for EMT cells), CD45 (red fluorescence for leukocytes), and Hoechst (blue fluorescence for nuclei). (A) Single CTC: CK+/Vimentin−/CD45− phenotype; (B) single CTC: CK+/Vimentin+/CD45− phenotype; (C) single CTC: CK−/Vimentin+/CD45− phenotype; (D) CTM: CK−/Vimentin+/CD45− phenotype; (E) CTM: CK+/Vimentin+/CD45− phenotype. The cells were analyzed at 40× magnification. Translational Oncology 2017 10, 431-441DOI: (10.1016/j.tranon.2017.02.007) Copyright © 2017 The Authors Terms and Conditions

Figure 3 (A) In 41 patients with stage IV gastric cancer, the PFS rate was significantly lower in patients with CTM than in those without CTM (P<.05). (B) In 41 patients with stage IV gastric cancer, the OS rate was significantly lower in patients with CTM than in those without CTM (P<.05). (C) In 30 patients with single CTCs or CTM, the PFS rate was significantly lower in patients with CTM than in those with only single CTCs (P<.05). (D) In 30 patients with single CTCs or CTM, the OS rate was significantly lower in patients with CTM than in those with only single CTCs (P<.05). Translational Oncology 2017 10, 431-441DOI: (10.1016/j.tranon.2017.02.007) Copyright © 2017 The Authors Terms and Conditions

Figure 4 Detection of CA125 expression in human gastric epithelial cell line GES-1 and human gastric cancer cell lines. (A) Immunofluorescent staining of GES-1, BGC823, MGC803, MKN45, and SGC7901 cell lines with CA125 (red fluorescence for CA125) and DAPI (blue fluorescence for nuclei). (B) Quantitative RT-PCR was carried out to detect relative CA125 mRNA expression in GES-1, BGC823, MGC803, MKN45, and SGC7901 cell lines. The cells were analyzed at 40× magnification. Translational Oncology 2017 10, 431-441DOI: (10.1016/j.tranon.2017.02.007) Copyright © 2017 The Authors Terms and Conditions