Journal: Clinical Cancer Research
Article Title: Clinical Significance for Risk Stratification of Papillary Thyroid Cancer by TUMORFISHER Circulating Tumor Cell Technique
doi: 10.1158/1078-0432.CCR-25-2694
Figure Lengend Snippet: In vitro validation of the Pep@MNP platform for specific capture and identification of thyroid cancer cells. A and B, Immunofluorescence staining of the capture marker EpCAM ( A ) and the identification marker CKmix ( B ) in thyroid cancer cell lines (BHT101 and BCPAP) and a negative control cell line (HL60). Nuclei were counterstained with DAPI. Scale bars, 10 μm. C, Quantitative analysis of the MFI of CKmix in BHT101, BCPAP, and HL60 cells, demonstrating significantly higher expression in thyroid cancer cells. D and E, FCM analysis confirming the surface expression of EpCAM ( D ) and high intracellular expression of CKmix ( E ) in BHT101 and BCPAP cells but not in HL60 cells. F, Scanning electron microscopy images showing the specific binding of Pep@MNPs to target BCPAP cells, whereas bare MNPs show minimal interaction. Both nanoparticle types show negligible binding to the negative control HL60 cells. Scale bars, 5 μm (top row) and 3 μm (bottom row). G, Representative immunofluorescence images defining the criteria for identifying a captured BCPAP cell (CTCs-like phenotype: DAPI+/CKmix+/CD45 − ) and distinguishing it from a co-captured white blood cell (WBC; DAPI+/CKmix − /CD45 + ). H, Capture sensitivity analysis. The graph shows the capture efficiency of the platform for varying numbers of BCPAP cells spiked into a solution. I, Capture specificity analysis. The graph compares the capture efficiency of the platform for cells with high EpCAM expression (BCPAP), low EpCAM expression (BHT101), and negative EpCAM expression (HL60). ****, P value < 0.0001.
Article Snippet: The human thyroid cancer cell lines BCPAP (RRID: CVCL_0153) and BHT101 (RRID: CVCL_1085), and the human promyelocytic leukemia cell line HL60 (RRID: CVCL_0002), were obtained from Procell Life Science & Technology in August 2021.
Techniques: In Vitro, Biomarker Discovery, Immunofluorescence, Staining, Marker, Negative Control, Expressing, Electron Microscopy, Binding Assay