Journal of Microscopy and Ultrastructure

ORIGINAL ARTICLE
Year
: 2019  |  Volume : 7  |  Issue : 3  |  Page : 136--140

In vivo bioluminescence-based monitoring of liver metastases from colorectal cancer: An experimental model


Paolo Magistri1, Cecilia Battistelli2, Gabriele Toietta3, Raffaele Strippoli2, Andrea Sagnotta1, Antonello Forgione4, Fabrizio Di Benedetto5, Stefania Uccini6, Paola Vittorioso7, Francesco D'Angelo1, Paolo Aurello1, Giovanni Ramacciato1, Giuseppe Nigri1 
1 Department of Medical and Surgical Sciences and Translational Medicine, Sapienza University of Rome, Rome, Italy
2 Department of Cellular Biotechnology and Hematology, Molecular Genetics Section, Sapienza University of Rome, Rome, Italy
3 Department of Research, Advanced Diagnostic and Technological Innovation – Translational Research Area, Regina Elena National Cancer Institute, Rome, Italy
4 Advanced International Mini-invasive Surgery (AIMS) Academy, Ospedale Niguarda Ca' Granda, Milan, Italy
5 Hepato-Pancreato-Biliary Surgery and Liver Transplantation Unit, University of Modena and Reggio Emilia, Modena, Italy
6 Department of Pathology, Sapienza University of Rome, Rome, Italy
7 Pasteur Institute Fondazione Cenci Bolognetti, Department of Biology and Biotechnology ‘Charles Darwin’, Sapienza University of Rome, Rome, Italy

Correspondence Address:
Prof. Giuseppe Nigri
Department of Medical and Surgical Sciences and Translational Medicine, Sapienza University of Rome, Via Di Grottarossa 1035 00189, Rome
Italy

Background: In this study we aimed to develop a new in vivo bioluminescence-based tool to monitor and to quantify colon cancer (CC) liver metastasis development. Methods: HCT 116 cells were transducted with pLenti6/V5-DEST-fLuc for constitutive expression of firefly luciferase. Infection was monitored analyzing endogenous bioluminescence using the IVIS Lumina II In vivo Imaging System and a positive clone constitutively expressing luciferase (HCT 116-fLuc) was isolated. HCT 116-fLuc cells were left untreated or treated with 1 μM GDC-0449, a Hedgehog pharmacological inhibitor. Moreover, 1 x 106 HCT 116-fLuc cells were implanted via intra-splenic injection in nude mice. Bioluminescence was analyzed in these mice every 7 days for 5 weeks. After that, mice were sacrificed and bioluminescence was analyzed on explanted livers. Results: We found that in vitro bioluminescence signal was significantly reduced when HCT 116-fLuc cells were treated with GDC-0449. Regarding in vivo data, bioluminescence sources consistent with hepatic anatomical localization were detected after 21 days from HCT 116-fLuc intrasplenic injection and progressively increased until the sacrifice. The presence of liver metastasis was further confirmed by ex-vivo bioluminescence analysis of explanted livers. Conclusions: Our in vitro results suggest that inhibition of Hedgehog pathway may hamper CC cell proliferation and impel for further studies. Regarding in vivo data, we set-up a strategy for liver metastasis visualization, that may allow follow-up and quantification of the entire metastatic process. This cost-effective technique would reduce experimental variability, as well as the number of sacrificed animals.


How to cite this article:
Magistri P, Battistelli C, Toietta G, Strippoli R, Sagnotta A, Forgione A, Di Benedetto F, Uccini S, Vittorioso P, D'Angelo F, Aurello P, Ramacciato G, Nigri G. In vivo bioluminescence-based monitoring of liver metastases from colorectal cancer: An experimental model.J Microsc Ultrastruct 2019;7:136-140


How to cite this URL:
Magistri P, Battistelli C, Toietta G, Strippoli R, Sagnotta A, Forgione A, Di Benedetto F, Uccini S, Vittorioso P, D'Angelo F, Aurello P, Ramacciato G, Nigri G. In vivo bioluminescence-based monitoring of liver metastases from colorectal cancer: An experimental model. J Microsc Ultrastruct [serial online] 2019 [cited 2019 Oct 18 ];7:136-140
Available from: http://www.jmau.org/article.asp?issn=2213-879X;year=2019;volume=7;issue=3;spage=136;epage=140;aulast=Magistri;type=0