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Blood, Vol. 112, Issue 6, 2489-2499, September 15, 2008
Molecular basis of bortezomib resistance: proteasome subunit β5 (PSMB5) gene mutation and overexpression of PSMB5 protein
Blood Oerlemans et al.
112: 2489
Supplemental materials for: Oerlemans et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 115 KB)
- Figure S1. Expression of Multidrug Resistance transporters in bortezomib sensitive- and resistant THP1 cells (PDF, 65.1 KB) -
Western blot analysis of MRP1-6, Pgp and BCRP in THP1/WT, THP/BTZ30 and THP1/BTZ200 cells. As positive controls served drug-selected or transfected cell lines as described by Scheffer et al.43 A representative picture of 2 separate experiments is depicted.
- Figure S2. Micro array analysis for resistance- and proteasome related genes in bortezomib-resistant THP1 cells (PDF, 32.4 KB) -
Ratios of gene expression levels in THP1/BTZ30, THP1/BTZ100 and THP1/BTZ(−100) cells relative to THP1/WT cells are depicted for selected genes implicated in drug metabolism, resistance, apoptosis and cell cycle regulation, and compared with those in the ubiquitin proteasome pathway. Experimental details are described in the Materials & Methods section.
- Figure S3. Expression of selected resistance, apoptosis and cell cycle proteins in THP1/WT cells and Bortezomib-resistant cells (PDF, 52 KB) -
Western blot analysis of hsp27, hsp90, apoptosis inhibitor protein XIAP and p21 (cyclin dependent kinase inhibitor 1A) in THP/WT cells and bortezomib-resistant THP1 cells. Note: induction of p21 in THP1/BTZ500 cells point to early growth inhibitory effects of 500 nM bortezomib in these cells. A representative picture of 2 separate experiments is shown.
- Figure S4. Intracellular proteasome β5 and α7 subunit distribution in THP1/WT and THP1/BTZ200 cells (PDF, 53.2 KB) -
Fluorescence microscopy of: (A)/(D); α7 proteasome subunits, (B)/E); β5 proteasome subunits and (C)/(F); merge of the α7 and β5 staining. Upper panels: THP1/WT cells; lower panels: THP1/BTZ200 cells.
- Figure S5. Upregulation of proteasome β5 subunit expression in bortezomib resistant CCRF-CEM cells (PDF, 51.3 KB) -
Human CCRF-CEM cells of T cells origin were chronically exposed to stepwise increasing concentrations of bortezomib over a period of 6 months. Expression levels of proteasome subunit β5 and α7 are depicted for THP1/WT cells and two selected bortezomib resistant variants grown at 7 nM bortezomib (BTZ7) and 200 nM bortezomib (BTZ200).
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