Chapter 14
Methods to Generate and Assay for Distinct Stages
of Cancer Metastasis in Adult Drosophila melanogaster
Jamie Adams, Andreu Casali, and Kyra Campbell
Abstract
Metastasis underlies the majority of cancer-related deaths. Until recently, research on this complex multistep process has been hindered by a lack of genetically tractable experimental models amenable to highthroughput analyses. This was recently overcome with the development of a model of metastatic colorectal
cancer (CRC) in adult flies, which relies on the activation of a partial-epithelial-to-mesenchymal transition
(EMT) in intestinal tumors. In this model, tumor cells are labeled with both GFP and luciferase reporters,
enabling high-throughput analyses. We report here the detailed protocol for generating the model, and
assaying for primary tumor burden and distinct stages of metastasis, including the number of circulating
tumor cells and secondary metastases.
Key words Cancer metastasis, Drosophila, Colorectal cancer
1 Introduction
Metastasis is the leading cause of cancer-related deaths globally. It is
a complex multistep process which involves the invasion of cells
from the primary tumor into surrounding tissues, migration and
dissemination through the body, and colonization of distant sites.
Invertebrate models such as Drosophila melanogaster (Drosophila)
are emerging as powerful tools to investigate the mechanisms
underlying malignancy and identify novel therapeutics [1]. While
there are many anatomical and physiological differences between
humans and flies, the number of cellular and organismic aspects of
tumorigenesis that they share is remarkable. Additionally, of particular relevance to modeling colorectal cancer (CRC), there are
striking similarities between the intestinal tracts of mammals and
of Drosophila [2]. Experimentally, Drosophila are highly amenable
to genetic manipulation, with a wealth of sophisticated genetic
tools and publically available reagents. Combined with their short
life-cycle, this allows for the analysis of large number of individuals
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_14,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
161
Methods to Generate and Assay for Distinct Stages
of Cancer Metastasis in Adult Drosophila melanogaster
Jamie Adams, Andreu Casali, and Kyra Campbell
Abstract
Metastasis underlies the majority of cancer-related deaths. Until recently, research on this complex multistep process has been hindered by a lack of genetically tractable experimental models amenable to highthroughput analyses. This was recently overcome with the development of a model of metastatic colorectal
cancer (CRC) in adult flies, which relies on the activation of a partial-epithelial-to-mesenchymal transition
(EMT) in intestinal tumors. In this model, tumor cells are labeled with both GFP and luciferase reporters,
enabling high-throughput analyses. We report here the detailed protocol for generating the model, and
assaying for primary tumor burden and distinct stages of metastasis, including the number of circulating
tumor cells and secondary metastases.
Key words Cancer metastasis, Drosophila, Colorectal cancer
1 Introduction
Metastasis is the leading cause of cancer-related deaths globally. It is
a complex multistep process which involves the invasion of cells
from the primary tumor into surrounding tissues, migration and
dissemination through the body, and colonization of distant sites.
Invertebrate models such as Drosophila melanogaster (Drosophila)
are emerging as powerful tools to investigate the mechanisms
underlying malignancy and identify novel therapeutics [1]. While
there are many anatomical and physiological differences between
humans and flies, the number of cellular and organismic aspects of
tumorigenesis that they share is remarkable. Additionally, of particular relevance to modeling colorectal cancer (CRC), there are
striking similarities between the intestinal tracts of mammals and
of Drosophila [2]. Experimentally, Drosophila are highly amenable
to genetic manipulation, with a wealth of sophisticated genetic
tools and publically available reagents. Combined with their short
life-cycle, this allows for the analysis of large number of individuals
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_14,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
161