A model for the detection of pancreatic ductal adenocarcinoma circulating tumor cells
Main Article Content
Keywords
pancreatic adenocarcinoma, circulating tumor cells, metastasis, bioluminescent imaging, orthotopic implantation
Abstract
Metastatic disease is the leading cause of pancreatic ductal adenocarcinoma (PDAC) associated death. PDAC cells invade and enter the bloodstream early, before frank malignancy can be detected. Our objective was to develop an in vivo assay enabling the identification and quantification of circulating tumor cells (CTCs) from primary orthotopic PDAC tumors. Human PDAC cells expressing luciferase and green fluorescent protein were orthotopically injected into the pancreas of mice utilizing ultrasound guidance. Bioluminescent imaging was conducted to identify and track tumor growth. CTCs were then isolated and analyzed by flow cytometry to detect GFP-expressing cancer cells. Tumor growth as measured by bioluminescent imaging increased over time. The concentration of CTCs correlated with the strength of bioluminescent imaging signal. In addition, livers bearing macroscopic disease were harvested for further imaging under fluorescence stereomicroscopy and confocal microscopy, which confirmed the presence of metastases. This study represents an orthotopic animal model that reliably detects the presence of CTCs from PDAC. There is a positive correlation between the concentrations of CTCs with overall tumor burden.
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References
1. Howlader N NA, Krapcho M, Miller D, Bishop K, Kosary CL, Yu M, Ruhl J, Tatalovich Z, Mariotto A, Lewis DR, Chen HS, Feuer EJ, Cronin KA (eds). (2017) SEER Cancer Statistics Review, 1975-2014, National Cancer Institute. United States SEER Program 1975-2014. National Cancer Institute, SEER Program. Bethesda, MD: NIH Pub.
2. Oettle H, Post S, Neuhaus P, Gellert K, Langrehr J, et al. (2007) Adjuvant chemotherapy with gemcitabine vs observation in patients undergoing curative-intent resection of pancreatic cancer: a randomized controlled trial. Jama 297: 267-277.
3. Asiyanbola B, Gleisner A, Herman JM, Choti MA, Wolfgang CL, et al. (2009) Determining Pattern of Recurrence Following Pancreaticoduodenectomy and Adjuvant 5-Flurouracil-Based Chemoradiation Therapy: Effect of Number of Metastatic Lymph Nodes and Lymph Node Ratio. Journal of Gastrointestinal Surgery 13: 752-759.
4. Neoptolemos JP, Stocken DD, Friess H, Bassi C, Dunn JA, et al. (2004) A Randomized Trial of Chemoradiotherapy and Chemotherapy after Resection of Pancreatic Cancer. New England Journal of Medicine 350: 1200-1210.
5. Rhim Andrew D, Mirek Emily T, Aiello Nicole M, Maitra A, Bailey Jennifer M, et al. (2012) EMT and Dissemination Precede Pancreatic Tumor Formation. Cell 148: 349-361.
6. Ashworth T (1869) A case of cancer in which cells similar to those in the tumours were seen in the blood after death. Australian Medical Journal 14: 146-147.
7. Alix-Panabieres C, Pantel K (2014) Challenges in circulating tumour cell research. Nat Rev Cancer 14: 623-631.
8. George TJ, Ogunwobi OO, Sheng W, Fan ZH, Liu C (2014) “Tissue is the Issue”: Circulating Tumor Cells in Pancreatic Cancer. Journal of gastrointestinal cancer 45: 222-225.
9. Bockhorn M, Jain RK, Munn LL (2007) Active versus passive mechanisms in metastasis: do cancer cells crawl into vessels, or are they pushed? The Lancet Oncology 8: 444-448.
10. Zheng X, Carstens JL, Kim J, Scheible M, Kaye J, et al. (2015) Epithelial-to-mesenchymal transition is dispensable for metastasis but induces chemoresistance in pancreatic cancer. Nature 527: 525-530.
11. Pantel K, Speicher MR (2016) The biology of circulating tumor cells. Oncogene 35: 1216-1224.
12. Labelle M, Hynes RO (2012) The initial hours of metastasis: the importance of cooperative host-tumor cell interactions during hematogenous dissemination. Cancer Discov 2: 1091-1099.
13. Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, et al. (2004) Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med 351: 781-791.
14. Cohen SJ, Punt CJ, Iannotti N, Saidman BH, Sabbath KD, et al. (2009) Prognostic significance of circulating tumor cells in patients with metastatic colorectal cancer. Ann Oncol 20: 1223-1229.
15. Scher HI, Jia X, de Bono JS, Fleisher M, Pienta KJ, et al. (2009) Circulating tumour cells as prognostic markers in progressive, castration-resistant prostate cancer: a reanalysis of IMMC38 trial data. Lancet Oncol 10: 233-239.
16. Naito T, Tanaka F, Ono A, Yoneda K, Takahashi T, et al. (2012) Prognostic impact of circulating tumor cells in patients with small cell lung cancer. J Thorac Oncol 7: 512-519.
17. Kulemann B, Rösch S, Seifert S, Timme S, Bronsert P, et al. (2017) Pancreatic cancer: Circulating Tumor Cells and Primary Tumors show Heterogeneous KRAS Mutations. Scientific Reports 7: 4510.
18. Ankeny JS, Court CM, Hou S, Li Q, Song M, et al. (2016) Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer. Br J Cancer 114: 1367-1375.
19. Poruk KE, Valero V, 3rd, Saunders T, Blackford AL, Griffin JF, et al. (2016) Circulating Tumor Cell Phenotype Predicts Recurrence and Survival in Pancreatic Adenocarcinoma. Ann Surg 264: 1073-1081.
20. Xie ZB, Yao L, Jin C, Fu DL (2016) Circulating tumor cells in pancreatic cancer patients: efficacy in diagnosis and value in prognosis. Discov Med 22: 121-128.
21. Kulemann B, Pitman MB, Liss AS, Valsangkar N, Fernandez-Del Castillo C, et al. (2015) Circulating tumor cells found in patients with localized and advanced pancreatic cancer. Pancreas 44: 547-550.
22. Han L, Chen W, Zhao Q (2013) Prognostic value of circulating tumor cells in patients with pancreatic cancer: A meta-analysis.
23. Muniz VP, Barnes JM, Paliwal S, Zhang X, Tang X, et al. (2011) The ARF tumor suppressor inhibits tumor cell colonization independent of p53 in a novel mouse model of pancreatic ductal adenocarcinoma metastasis. Molecular cancer research : MCR 9: 867-877.
24. Du J, Cieslak JA, 3rd, Welsh JL, Sibenaller ZA, Allen BG, et al. (2015) Pharmacological Ascorbate Radiosensitizes Pancreatic Cancer. Cancer Res 75: 3314-3326.
25. Oettle H, Post S, Neuhaus P, et al. (2007) Adjuvant chemotherapy with gemcitabine vs observation in patients undergoing curative-intent resection of pancreatic cancer: A randomized controlled trial. JAMA 297: 267-277.
26. Maheswaran S, Haber DA (2010) Circulating tumor cells: a window into cancer biology and metastasis. Current Opinion in Genetics & Development 20: 96-99.
27. Haeno H, Gonen M, Davis MB, Herman JM, Iacobuzio-Donahue CA, et al. (2012) Computational modeling of pancreatic cancer reveals kinetics of metastasis suggesting optimum treatment strategies. Cell 148: 362-375.
1. Howlader N NA, Krapcho M, Miller D, Bishop K, Kosary CL, Yu M, Ruhl J, Tatalovich Z, Mariotto A, Lewis DR, Chen HS, Feuer EJ, Cronin KA (eds). (2017) SEER Cancer Statistics Review, 1975-2014, National Cancer Institute. United States SEER Program 1975-2014. National Cancer Institute, SEER Program. Bethesda, MD: NIH Pub.
2. Oettle H, Post S, Neuhaus P, Gellert K, Langrehr J, et al. (2007) Adjuvant chemotherapy with gemcitabine vs observation in patients undergoing curative-intent resection of pancreatic cancer: a randomized controlled trial. Jama 297: 267-277.
3. Asiyanbola B, Gleisner A, Herman JM, Choti MA, Wolfgang CL, et al. (2009) Determining Pattern of Recurrence Following Pancreaticoduodenectomy and Adjuvant 5-Flurouracil-Based Chemoradiation Therapy: Effect of Number of Metastatic Lymph Nodes and Lymph Node Ratio. Journal of Gastrointestinal Surgery 13: 752-759.
4. Neoptolemos JP, Stocken DD, Friess H, Bassi C, Dunn JA, et al. (2004) A Randomized Trial of Chemoradiotherapy and Chemotherapy after Resection of Pancreatic Cancer. New England Journal of Medicine 350: 1200-1210.
5. Rhim Andrew D, Mirek Emily T, Aiello Nicole M, Maitra A, Bailey Jennifer M, et al. (2012) EMT and Dissemination Precede Pancreatic Tumor Formation. Cell 148: 349-361.
6. Ashworth T (1869) A case of cancer in which cells similar to those in the tumours were seen in the blood after death. Australian Medical Journal 14: 146-147.
7. Alix-Panabieres C, Pantel K (2014) Challenges in circulating tumour cell research. Nat Rev Cancer 14: 623-631.
8. George TJ, Ogunwobi OO, Sheng W, Fan ZH, Liu C (2014) “Tissue is the Issue”: Circulating Tumor Cells in Pancreatic Cancer. Journal of gastrointestinal cancer 45: 222-225.
9. Bockhorn M, Jain RK, Munn LL (2007) Active versus passive mechanisms in metastasis: do cancer cells crawl into vessels, or are they pushed? The Lancet Oncology 8: 444-448.
10. Zheng X, Carstens JL, Kim J, Scheible M, Kaye J, et al. (2015) Epithelial-to-mesenchymal transition is dispensable for metastasis but induces chemoresistance in pancreatic cancer. Nature 527: 525-530.
11. Pantel K, Speicher MR (2016) The biology of circulating tumor cells. Oncogene 35: 1216-1224.
12. Labelle M, Hynes RO (2012) The initial hours of metastasis: the importance of cooperative host-tumor cell interactions during hematogenous dissemination. Cancer Discov 2: 1091-1099.
13. Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, et al. (2004) Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med 351: 781-791.
14. Cohen SJ, Punt CJ, Iannotti N, Saidman BH, Sabbath KD, et al. (2009) Prognostic significance of circulating tumor cells in patients with metastatic colorectal cancer. Ann Oncol 20: 1223-1229.
15. Scher HI, Jia X, de Bono JS, Fleisher M, Pienta KJ, et al. (2009) Circulating tumour cells as prognostic markers in progressive, castration-resistant prostate cancer: a reanalysis of IMMC38 trial data. Lancet Oncol 10: 233-239.
16. Naito T, Tanaka F, Ono A, Yoneda K, Takahashi T, et al. (2012) Prognostic impact of circulating tumor cells in patients with small cell lung cancer. J Thorac Oncol 7: 512-519.
17. Kulemann B, Rösch S, Seifert S, Timme S, Bronsert P, et al. (2017) Pancreatic cancer: Circulating Tumor Cells and Primary Tumors show Heterogeneous KRAS Mutations. Scientific Reports 7: 4510.
18. Ankeny JS, Court CM, Hou S, Li Q, Song M, et al. (2016) Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer. Br J Cancer 114: 1367-1375.
19. Poruk KE, Valero V, 3rd, Saunders T, Blackford AL, Griffin JF, et al. (2016) Circulating Tumor Cell Phenotype Predicts Recurrence and Survival in Pancreatic Adenocarcinoma. Ann Surg 264: 1073-1081.
20. Xie ZB, Yao L, Jin C, Fu DL (2016) Circulating tumor cells in pancreatic cancer patients: efficacy in diagnosis and value in prognosis. Discov Med 22: 121-128.
21. Kulemann B, Pitman MB, Liss AS, Valsangkar N, Fernandez-Del Castillo C, et al. (2015) Circulating tumor cells found in patients with localized and advanced pancreatic cancer. Pancreas 44: 547-550.
22. Han L, Chen W, Zhao Q (2013) Prognostic value of circulating tumor cells in patients with pancreatic cancer: A meta-analysis.
23. Muniz VP, Barnes JM, Paliwal S, Zhang X, Tang X, et al. (2011) The ARF tumor suppressor inhibits tumor cell colonization independent of p53 in a novel mouse model of pancreatic ductal adenocarcinoma metastasis. Molecular cancer research : MCR 9: 867-877.
24. Du J, Cieslak JA, 3rd, Welsh JL, Sibenaller ZA, Allen BG, et al. (2015) Pharmacological Ascorbate Radiosensitizes Pancreatic Cancer. Cancer Res 75: 3314-3326.
25. Oettle H, Post S, Neuhaus P, et al. (2007) Adjuvant chemotherapy with gemcitabine vs observation in patients undergoing curative-intent resection of pancreatic cancer: A randomized controlled trial. JAMA 297: 267-277.
26. Maheswaran S, Haber DA (2010) Circulating tumor cells: a window into cancer biology and metastasis. Current Opinion in Genetics & Development 20: 96-99.
27. Haeno H, Gonen M, Davis MB, Herman JM, Iacobuzio-Donahue CA, et al. (2012) Computational modeling of pancreatic cancer reveals kinetics of metastasis suggesting optimum treatment strategies. Cell 148: 362-375.
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Published
Sep 5, 2018
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Alexander MS, O’Leary BR, Moose D, Du J, Henry MD, Cullen JJ. A model for the detection of pancreatic ductal adenocarcinoma circulating tumor cells. J Biol Methods [Internet]. 2018 Sep. 5 [cited 2024 Apr. 25];5(3):e97. Available from: https://jbmethods.org/index.php/jbm/article/view/250
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