Abstract
Background:Anti-apoptotic BCL-2 family members antagonise apoptosis by sequestering their pro-apoptotic counterparts. The balance between the different BCL-2 family members forms the basis of BH3 profiling, a peptide-based technique used to predict chemosensitivity of cancer cells. Recent identification of cell-permeable, selective inhibitors of BCL-2, BCL-XL and MCL-1, further facilitates the determination of the BCL-2 family dependency of cancer cells.Methods:We use BH3 profiling in combination with cell death analyses using a chemical inhibitor toolkit to assess chemosensitivity of cancer cells.Results:Both BH3 profiling and the inhibitor toolkit effectively predict chemosensitivity of cells addicted to a single anti-apoptotic protein but a combination of both techniques is more instructive when cell survival depends on more than one anti-apoptotic protein.Conclusions:The inhibitor toolkit provides a rapid, inexpensive and simple means to assess the chemosensitivity of tumour cells and in conjunction with BH3 profiling offers much potential in personalising cancer therapy.
Highlights
Anti-apoptotic BCL-2 family members antagonise apoptosis by sequestering their pro-apoptotic counterparts
A-1331852 and A-1210477 have been identified as specific inhibitors of BCL-XL and MCL-1, respectively (Leverson et al, 2015a, b)
To validate the efficacy of BH3 profiling on cells addicted to specific BCL-2 family members, primary chronic lymphocytic leukaemia (CLL) cells, addicted to BCL-2 (Del Gaizo Moore et al, 2007; Vogler et al, 2009a, b), MOLT-4 and H929 cell lines, addicted to BCL-XL (Leverson et al, 2015a) and MCL-1 (Leverson et al, 2015b), respectively, and H1299 cells addicted to both BCL-XL and MCL-1 (Varadarajan et al, 2013) were selected
Summary
We use BH3 profiling in combination with cell death analyses using a chemical inhibitor toolkit to assess chemosensitivity of cancer cells. Peripheral blood samples from CLL patients were obtained with patient consent and local ethics committee approval. BRITISH JOURNAL OF CANCER and cultured as described (Vogler et al, 2009b). MOLT-4 and H1299, an AML and non-small cell lung carcinoma cell line, respectively, were cultured in RPMI 1640 medium supplemented with 10% foetal calf serum and 5 mM L-glutamine (Life Technologies Inc., Paisley, UK). H929, a multiple myeloma cell line, was cultured in the same medium supplemented with 0.02% 2-mercaptoethanol. All cell lines were from ATCC (Middlesex, UK). ABT-199, A-1331852 and A-1210477 were kindly supplied by Abbvie Inc., (North Chicago, IL, USA).
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