American Association for Cancer Research
15417786mcr160011-sup-160749_1_supp_3355868_y2g93p.pptx (41.36 MB)

Supplemental Figure 1 from Development of a Novel c-MET–Based CTC Detection Platform

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posted on 2023-04-03, 17:10 authored by Tian Zhang, Rengasamy Boominathan, Brad Foulk, Chandra Rao, Gabor Kemeny, John H. Strickler, James L. Abbruzzese, Michael R. Harrison, David S. Hsu, Patrick Healy, Jing Li, Cinthia Pi, Katherine M. Prendergast, Carey Hobbs, Sarah Gemberling, Daniel J. George, Herbert I. Hurwitz, Mark Connelly, Mariano A. Garcia-Blanco, Andrew J. Armstrong

Immunofluorescence characterization of EpCAM and c-MET in cell lines. Immunofluorescence of SNU5, HeLa and SIHA cell lines, with (clockwise from top left of each panel) external c-MET, EpCAM, DAPI from EpCAM slide, and DAPI from c-MET slide.



Amplification of the MET oncogene is associated with poor prognosis, metastatic dissemination, and drug resistance in many malignancies. We developed a method to capture and characterize circulating tumor cells (CTC) expressing c-MET using a ferromagnetic antibody. Immunofluorescence was used to characterize cells for c-MET, DAPI, and pan-CK, excluding CD45+ leukocytes. The assay was validated using appropriate cell line controls spiked into peripheral blood collected from healthy volunteers (HV). In addition, peripheral blood was analyzed from patients with metastatic gastric, pancreatic, colorectal, bladder, renal, or prostate cancers. CTCs captured by c-MET were enumerated, and DNA FISH for MET amplification was performed. The approach was highly sensitive (80%) for MET-amplified cells, sensitive (40%–80%) for c-MET–overexpressed cells, and specific (100%) for both c-MET–negative cells and in 20 HVs. Of 52 patients with metastatic carcinomas tested, c-MET CTCs were captured in replicate samples from 3 patients [gastric, colorectal, and renal cell carcinoma (RCC)] with 6% prevalence. CTC FISH demonstrated that MET amplification in both gastric and colorectal cancer patients and trisomy 7 with gain of MET gene copies in the RCC patient. The c-MET CTC assay is a rapid, noninvasive, sensitive, and specific method for detecting MET-amplified tumor cells. CTCs with MET amplification can be detected in patients with gastric, colorectal, and renal cancers.Implications: This study developed a novel c-MET CTC assay for detecting c-MET CTCs in patients with MET amplification and warrants further investigation to determine its clinical applicability. Mol Cancer Res; 14(6); 539–47. ©2016 AACR.

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