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Fluorescence‐based experimental model to evaluate the concomitant effect of drugs on the tumour microenvironment and cancer cells
Author(s) -
Ramasamy Karthik,
Khatun Hazera,
Macpherson Lee,
Caley Mathew P.,
Sturge Justin,
Mufti Ghulam J.,
Schey Stephen A.,
Calle Yolanda
Publication year - 2012
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.2012.09103.x
Subject(s) - stromal cell , cancer cell , chemistry , cancer research , bone marrow , tumor microenvironment , bortezomib , bisphosphonate , bone resorption , multiple myeloma , fibroblast , microbiology and biotechnology , cancer , pathology , biology , medicine , in vitro , immunology , biochemistry , osteoporosis , tumor cells
Summary The response of the tumour microenvironment to anti‐cancer drugs can influence treatment efficacy. Current drug‐screening methodologies fail to distinguish and quantify simultaneously the concomitant effect of drugs on the tumour stroma and cancer cells. To overcome this limitation we have developed a fluorescence‐based experimental model that employs m C herry‐labelled stromal cells (e.g. bone marrow fibroblastic stromal cells) co‐cultured in direct contact with enhanced green fluorescent protein‐labelled tumour cell lines for accurate assessment of proliferation and viability in both cell compartments and adhesion of tumour cells. Additionally, we used fluorescence‐based image analysis to determine morphological changes that correlate with cell function (e.g. morphology of the actin cytoskeleton and nuclearity of osteoclasts to predict their bone resorption activity). Using this platform we have revealed that dexamethasone induces HS 5 fibroblast proliferation and contact with multiple myeloma cells via a process involving S rc/c‐ A bl kinases. Osteoclasts also inhibited dexamethasone‐induced apoptosis in myeloma cells while retaining their normal morphology and functionality in bone resorption. Myeloma resistance to dexamethasone mediated by HS 5 cells and osteoclasts was reversed by treatment with the S rc/c‐ A bl inhibitor dasatinib but not with bortezomib. This new experimental platform provides a more precise screening of new therapeutics for improved efficacy of tumour cell killing within the bone marrow microenvironment.