ABSTRACT
Title
Generation of Small Cell Lung Cancer(SCLC) bioluminescent xenograft orthotopic mouse models and evaluation of anti-tumoral activity of different chemoterapic agents.
Authors
M. Ruocco
Doctorate School in Pharmacology and Toxicology
Dept. of Pharmacology – University of Bologna, Italy
Doctorate School in Pharmacology and Toxicology
Dept. of Pharmacology – University of Bologna, Italy
Abstract
Each year approximately 1.4 million people worldwide are diagnosed with lung cancer. Small cell lung cancer (SCLC) is initially chemosensitive, but rapidly relapses in a chemoresistant form with an overall survival of <5%. Consequently, novel therapies are urgently required. The purpose of this work is to develop a preclinical mouse model of SCLC to further investigate the mechanisms involved in the neoplasia and to evaluate new therapeutic agents[1].
The mouse model was created by inoculation of SCLC H69 human cell line. The tumor onset and progression were evaluated with a bioluminescence imaging (BLI) technique based on luciferase reporter gene extracted from the North American firefly (Photinus pyralis). In this regard, before being grafted in mouse lung, the tumor cells were transfected to permanently express luciferase gene. Afterwards, mice were monitored every seven days by a CCD camera able to detect photons emission after intraperitoneal injection of D-Luciferin [2], [3].
The mouse model has shown an incidence of a 100% and a 2 weeks latency. Mice were monitored for three months with an endpoint at 70 days from cells implantation.
The BLI showed the presence of bone metastasis in mice, reflecting the behaviour of human SCLC evolution.
To validate the mouse model as an instrument to evaluate the therapy response using BLI, we used Cisplatin (CDDP) and Etoposide (VP-16) in combination, since they are considered the first line therapy in clinic [4]. After 4 weeks of drug administration, the mice showed a decrease of BLI signal and a prolonged survival with an endpoint at 85 days from cells implantation.
At the end of the study, a histological characterization of the tumor has been performed.
We created a new xenograft orthotopic BLI mouse model of SCLC, that showed a tumor evolution, reflecting the human clinical course of this tumor, including the presence of bone metastasis. In vivo molecular BLI allowed the non-invasive, fast and accurate monitoring of tumor evolution, since the first phases of tumor development. Finally, the BLI demonstrated to be a suitable mean to evaluate new drugs in preclinical studies for the treatment of this highly aggressive cancer.
1) Pardo O. et al.(2009) – Cancer Res.Nov 15;69(22):8645-51
2) Alnawaz R et al.(2000) - Neoplasia;491-495:2(6).
3) Ruxana T et al.(2005) - Proc Am Thorac Soc; (2)..
4) Puglisi M et al.(2010) - British Journal of Cancer 102, 629–638
The mouse model was created by inoculation of SCLC H69 human cell line. The tumor onset and progression were evaluated with a bioluminescence imaging (BLI) technique based on luciferase reporter gene extracted from the North American firefly (Photinus pyralis). In this regard, before being grafted in mouse lung, the tumor cells were transfected to permanently express luciferase gene. Afterwards, mice were monitored every seven days by a CCD camera able to detect photons emission after intraperitoneal injection of D-Luciferin [2], [3].
The mouse model has shown an incidence of a 100% and a 2 weeks latency. Mice were monitored for three months with an endpoint at 70 days from cells implantation.
The BLI showed the presence of bone metastasis in mice, reflecting the behaviour of human SCLC evolution.
To validate the mouse model as an instrument to evaluate the therapy response using BLI, we used Cisplatin (CDDP) and Etoposide (VP-16) in combination, since they are considered the first line therapy in clinic [4]. After 4 weeks of drug administration, the mice showed a decrease of BLI signal and a prolonged survival with an endpoint at 85 days from cells implantation.
At the end of the study, a histological characterization of the tumor has been performed.
We created a new xenograft orthotopic BLI mouse model of SCLC, that showed a tumor evolution, reflecting the human clinical course of this tumor, including the presence of bone metastasis. In vivo molecular BLI allowed the non-invasive, fast and accurate monitoring of tumor evolution, since the first phases of tumor development. Finally, the BLI demonstrated to be a suitable mean to evaluate new drugs in preclinical studies for the treatment of this highly aggressive cancer.
1) Pardo O. et al.(2009) – Cancer Res.Nov 15;69(22):8645-51
2) Alnawaz R et al.(2000) - Neoplasia;491-495:2(6).
3) Ruxana T et al.(2005) - Proc Am Thorac Soc; (2)..
4) Puglisi M et al.(2010) - British Journal of Cancer 102, 629–638