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. 2020 Oct 19;10:17649. doi: 10.1038/s41598-020-74806-2

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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P-AscH⁻ slows PDAC tumor growth, reduces ascites development and decreases CTCs in vivo. (A) The growth of the pancreatic tumors was tracked weekly using bioluminescent imaging. Pancreatic tumors were formed through ultrasound guided orthotopic injections of pancreatic tumors cells into the pancreas of nude mice. The representative images show tumor burden between saline treated mice and P-AscH⁻ treated mice on day 49 of the experiment. (B) P-AscH⁻ significantly decreased tumor growth over time as determined by bioluminescent imaging. Treatments of P-AscH⁻ or saline were started on day 35 following tumor cell injections. A region of interest was placed around each mouse image and total photon flux (photons per second) was quantified A significant decrease in tumor growth rate was seen after initiation of therapy as indicated by bioluminescent signal strength. Data represent the mean photon flux compared to controls ± SE (n = 6–10, *p < 0.05; one-way ANOVA with Bonferroni’s multiple comparisons). (C) Saline treated mice were found to have significantly increased CTCs per tumor burden as measured by photon flux compared to P-AscH⁻ treated mice (2.4 × 10^–6 CTCs/photon flux vs 9.3 × 10^–8 CTCs/photon flux). Data represent the mean of CTCs/photon flux ± SE (n = 6–10, *p < 0.05; Mann–Whitney). (D) P-AscH⁻ decreased the number of CTCs. Saline treated mice were found to have increased CTCs/mL compared to P-AscH⁻ treated mice (3,460 +/− 1232 CTCs/mL vs 80 +/− 50 CTCs/mL) Data represent the mean of CTCs/mL ± SE (n = 6–10, *p < 0.05; Mann–Whitney). (E) The number of CTCs was increased in mice with metastatic disease. CTCs were isolated on day 50 of the experiment following a cardiac blood draw. Mice with localized tumors were found to have fewer CTCs/mL than mice with evidence of metastatic disease (900 +/− 505 CTCs/mL vs 3,590 +/− 1570 CTCs/mL). Data represent the mean of CTCs/mL ± SE (n = 8, *p < 0.05; Mann–Whitney). (F) Mice were evaluated daily for ascites accumulation following pancreatic injection of tumor cells. P-AscH⁻ decreased the percentage of mice with ascites (n = 8, *p < 0.05) compared to control (log-rank).