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Plastin-3 Transformed Circulating Tumor Cells in Hepatic Portal Vein of Pancreatic Cancer Patient for Predicting Liver Metastasis Application Note Introduction ● Pancreatic cancer is an asymptomatic disease which is usually diagnosed during advanced stage with high incidence of liver metastasis1. ● Evidence of liver metastasis is a principal determinant of disease management and clinical outcome in pancreatic cancer patients2. ● Whipple operation is a beneficial surgical procedure for localized pancreatic cancer, whereas palliative procedures are recommended for metastatic disease. ● Plastin-3, a metastasis-specific gene located on chromosome Xq23, is an epithelial mesenchymal transition (EMT) marker associated with colorectal and breast cancers3. ● Presence of Plastin-3 transformed circulating tumor cells (CTCs) in hepatic portal vein could serve as a clinical predictor of liver metastasis4. ● We applied CytoBot™ negative enrichment system, EpCAM, PanCK, and Plastin-3 monoclonal antibodies to identify epithelial CTCs in hepatic portal vein of a pancreatic cancer patient. Materials & Methods ● Portal vein blood of pancreatic cancer patient was collected in ACD Tube (364606, BD). ● The ACD Tube contained 7.5 mL blood was loaded into the CytoBot™ System and CTCs were applied on Abnova Adhesive Slide (U0320, Abnova) after the negative enrichment was completed automatically by CytoBot™ System. ● Immunofluorescence staining for detecting CTCs was performed using PanCK, Plastin-3, CD45 (KA4585, Abnova), DAPI as the instruction of protocol. ● Imaging was performed using Nikon Eclipse Ti-E fluorescent inverted microscope. Results ● CTC Counts: In 7.5 mL blood of pancreatic cancer patient, 5 cells count as CTC (PanCK+, Plastin-3+, CD45-, DAPI+). Merged PanCK CD45 Plastin-3 Nucleus Figure 1. Representative images of CTC (white arrow) and WBCs from pancreatic cancer patient. CTC was detected by using immunofluorescence staining for PanCK (FITC, green), Plastin-3 (Alexa647, red), CD45 (PE, orange) and Nucleus (DAPI, blue). Discussions ● Effective clinical management of pancreatic cancer patients depends on a complete medical workup to determine exact nature of metastasis. ● A clinical predictor of liver metastasis would be highly desirable before Whipple procedure for “localized” disease to assess the probability of metastasis before surgery. ● Access to the hepatic portal vein for sampling and detection of EMT-transformed circulating tumor cells would be a valuable adjunct to the clinical management. ● A combination of PanCK and Plastin-3 monoclonal antibodies successfully identify circulating pancreatic cancer cells using an antibody-based, negative enrichment. ● This case study is the first report of Plastin-3 EMT expression in pancreatic CTCs identified with EpCAM and PanCK epithelial cell markers. ● A larger cohort study will provide clinically evidence for establishing Plastin-3 transformed epithelial CTCs as a predictor of liver metastasis in “localized” pancreatic cancer patients. www.abnova.com Abnova Corporation 9F, No. 108, Jhouzih St., Neihu, Taipei 114, Taiwan Tel: + 886 2 8751 1888 Fax: + 886 2 6602 1218 References 1. Beger HG, Rau B, Gansauge F, Poch B, Link KH. Treatment of pancreatic cancer: challenge of the facts. World journal of surgery, 2003, 27(10):1075-84 2. Li D, Xie K, Wolff R, Abbruzzese JL. Pancreatic cancer. Lancet, 2004, 363(9414):1049-57 3. Yokobori T, et al. Plastin3 is a novel marker for circulating tumor cells undergoing the epithelial-mesenchymal transition and is associated with colorectal cancer prognosis. Cancer research, 2013, 73(7):2059-69 4. Lyberopoulou A, et al. Mutational analysis of circulating tumor cells from colorectal cancer patients and correlation with primary tumor tissue. PloS one, 2015, 10(4):e0123902 www.abnova.com Abnova Corporation 9F, No. 108, Jhouzih St., Neihu, Taipei 114, Taiwan Tel: + 886 2 8751 1888 Fax: + 886 2 6602 1218