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In ALL cell lines, ABT-263 was found to synergize with several different anti-neoplastic agents including daunorubicin, bortezomib, apicidin, and the multi-tyrosine inhibitor sunitinib (34)

In ALL cell lines, ABT-263 was found to synergize with several different anti-neoplastic agents including daunorubicin, bortezomib, apicidin, and the multi-tyrosine inhibitor sunitinib (34). exhibited high sensitivity to Bcl-2 inhibition by the BH3 mimetic compounds. Overall our results showed that primary ALL cultures were both more sensitive to BH3 mimetics and more uniform in their response than established ALL cell lines which have been evaluated previously. Further, the primary cell Crotonoside model characterized here offers a powerful system for preclinical testing of novel drugs and drug combinations to treat ALL. activity in a wide range of cancer cell lines, primary leukemia cells, and xenograft models (10-17). Additionally, Phase I and II clinical trials conducted for several types of cancer have shown promising results (13, 14, 18-20). Because a limitation of ABT-263 is thrombocytopenia due to Bcl-xL inhibition in circulating platelets, the derivative ABT-199 was recently developed, which is selective for Bcl-2 and exhibits anti-tumor activity without significant thrombocytopenia (21). Acute lymphoblastic leukemia (ALL) affects both adults and children (22, 23). Because cure rates have begun to plateau, new classes of therapeutic agents are needed, but these are difficult to evaluate systematically in patients especially in the context of polychemotherapy. Many continuously proliferating ALL cell lines have been established (24, 25), but after extensive propagation they have likely acquired properties which deviate from the originating primary cells. This emphasizes the need for preclinical cell models of ALL that more closely represent the disease. Recently, conditions were established for the expansion and long-term culture of primary adult ALL cells using a defined media that lacked Crotonoside serum and hematopoietic growth factors (26). This system provides a unique and powerful tool for the preclinical evaluation of novel therapies for ALL. In the present study, we examined ABT-263 and ABT-199 sensitivity, and Bcl-2 dependence and function, in several of these ALL cultures as well as in freshly isolated pediatric ALL blasts. These results demonstrate the utility of these expanded primary cultures for preclinical studies of ALL, provide mechanistic insight into the determinants of level of sensitivity and resistance to BH3 mimetics, and have important implications for the optimal use of these compounds in adult and pediatric ALL. Materials and Methods Materials, Cell extraction and immunoblotting, Caspase-3 assay, and Co-immunoprecipiation observe Supplementary Materials. Cell tradition KB3 cells (HeLa subline) were managed in DMEM, and RS4;11 and NALM-6 cell lines were taken care of in RPMI-1640 medium, supplemented with Crotonoside 10% bovine growth serum, 2 mM L-glutamine, 50 devices/mL penicillin, and 50 g/mL streptomycin. ALL cell cultures were maintained in suspension as explained (26) in Iscoves revised Dulbeccos medium (IMDM) comprising serum-free product (10 g/mL cholesterol, 6 mg/mL human being serum albumin, 0.5 g/mL amphotericin, 1 g/mL insulin, 200 g/mL human apo-transferrin, 50 M 2-mercaptoethanol, 2 mM glutamine and 50 units/mL penicillin). Mcl-1-dependent and Bcl-2-dependent leukemia cell lines were explained previously (27). Cells were managed at 37C and 5% CO2. Authentication of the cell lines and ALL cultures was founded via short tandem repeat (STR) profiling in September, 2014, by Genetica DNA Laboratories (LabCorp Speciality Screening Group, Burlington, NC). The STR profile of each cell line matched that of research profiles available in the ATCC database. The primary ALL cell tradition profiles did not match any repository cell lines, as expected, and each profile was unique with respect to the others. Rabbit polyclonal to PIWIL2 Cell viability assay Cell Crotonoside viability was identified using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium.