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Updated: Aug 14, 2025
Flow Cytometry-based Assay for the Monitoring of NK Cell Functions
Published on: October 30, 2016
Study of the effects of NK-tumor cell interaction by proteomic analysis and imaging
Chiara Lavarello1, Paola Orecchia2, Andrea Petretto1
1Core Facilities-Clinical Proteomics and Metabolomics, IRCCS Istituto Giannina Gaslini, Genoa, Italy.
Abstract:
Natural Killer (NK) cells play a pivotal role in the elimination of tumor cells. The interactions that NK cells can establish with cancer cells in the tumor microenvironment (TME) are crucial for the outcome of the anti-tumor response, possibly resulting in mechanisms able to modulate NK cell effector functions on the one side, and to modify tumor cell phenotype and properties on the other side. This chapter will describe two different experimental approaches for the evaluation of NK-tumor cell interactions. First, a detailed protocol for the setting up of NK-tumor cell co-cultures will be illustrated, followed by information on cell imaging techniques, useful for assessing cell morphology and cytoskeletal changes. The second part will be focused on the description of a proteomic approach aimed at investigating the effect of this crosstalk from another point of view, i.e., characterizing the cellular and molecular pathways modulated in tumor cells following interaction with NK cells. The chapter centers on the interaction between NK and melanoma cells and refers to experimental approaches we set up to study the effects of this cross-talk on the process of the Epithelial-to-Mesenchymal Transition (EMT). Nevertheless, the described protocols can be quite easily adapted to study the interaction of NK cells with adherent tumor cell lines of different origin and histotype, as in our original study, we also analyzed possible NK-induced morphologic changes in the cervix adenocarcinoma HeLa cells and the colon cancer HT29 cells.
Insights
Natural Killer (NK) cells combat cancer by interacting with tumor cells. This study details methods to analyze these interactions, focusing on melanoma and their impact on Epithelial-to-Mesenchymal Transition (EMT).
Area of Science:
- Immunology
- Cell Biology
- Cancer Research
Background:
- Natural Killer (NK) cells are crucial for anti-tumor immunity.
- NK cell interactions within the tumor microenvironment (TME) influence both NK cell function and tumor cell phenotype.
- Understanding NK-tumor cell crosstalk is vital for developing effective cancer therapies.
Purpose of the Study:
- To describe experimental approaches for evaluating NK-tumor cell interactions.
- To investigate the impact of NK cell interactions on tumor cell phenotype, specifically focusing on Epithelial-to-Mesenchymal Transition (EMT).
- To provide adaptable protocols for studying NK cell interactions with various cancer cell lines.
Main Methods:
- Detailed protocols for setting up NK-tumor cell co-cultures.
- Cell imaging techniques to assess morphology and cytoskeletal changes.
- Proteomic approaches to characterize molecular pathways modulated in tumor cells post-interaction.
Main Results:
- The study presents methods applicable to NK-melanoma cell interactions, with potential for adaptation to other cancer types (e.g., HeLa, HT29).
- Evaluated NK-induced morphologic changes and molecular pathway modulation in tumor cells.
- Assessed the effects of NK-tumor cell crosstalk on the Epithelial-to-Mesenchymal Transition (EMT) process.
Conclusions:
- The described co-culture and proteomic methods provide valuable tools for dissecting NK-tumor cell interactions.
- These approaches can elucidate how NK cells influence tumor cell behavior, including EMT.
- The protocols are versatile and can be applied to diverse cancer cell types for broader research applications.
