OH-2, a hyperdiploid myeloma cell line without an IGH translocation, has a complex translocation juxtaposing MYC near MAFB and the IGK locus

Våtsveen, Thea Kristin; Tian, Erming; Kresse, Stine H.; Meza‐Zepeda, Leonardo A.; Gabrea, Ana; Glebov, O K; Dai, Hong; Sundan, Anders; Kuehl, W. Michael; Børset, Magne

doi:10.1016/j.leukres.2009.03.001

Cited by 16 publications

(23 citation statements)

References 22 publications

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“…The myeloma cell lines used in this study were ANBL-6 (a kind gift from Dr Diane Jelinek, Mayo Clinic, Rochester, MN, USA), INA-6 (a kind gift from Dr Martin Gramatzki, Erlangen, Germany 25 ), U266 (ATCC, Rockville, MD, USA), IH-1, 26 OH-2, 27 JJN3 (a kind gift from Dr Jennifer Ball, Department of Immunology, University of Birmingham, UK) and KJON (established in our laboratory). ANBL-6 and INA-6 cells were grown in 10% heat-inactivated fetal calf serum in RPMI-1640 (hereafter described as RPMI, Sigma-Aldrich, St. Louis, MO, USA, R8758) supplemented with recombinant human interleukin (IL)-6 (1 ng/ml) (Gibco, Life Technologies/Thermo Fisher, Waltham, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

Intracellular glutathione determines bortezomib cytotoxicity in multiple myeloma cells

Starheim

Holien

Misund

et al. 2016

Blood Cancer Journal

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Multiple myeloma (myeloma in short) is an incurable cancer of antibody-producing plasma cells that comprise 13% of all hematological malignancies. The proteasome inhibitor bortezomib has improved treatment significantly, but inherent and acquired resistance to the drug remains a problem. We here show that bortezomib-induced cytotoxicity was completely dampened when cells were supplemented with cysteine or its derivative, glutathione (GSH) in ANBL-6 and INA-6 myeloma cell lines. GSH is a major component of the antioxidative defense in eukaryotic cells. Increasing intracellular GSH levels fully abolished bortezomib-induced cytotoxicity and transcriptional changes. Elevated intracellular GSH levels blocked bortezomib-induced nuclear factor erythroid 2-related factor 2 (NFE2L2, NRF2)-associated stress responses, including upregulation of the xCT subunit of the Xc- cystine-glutamate antiporter. INA-6 cells conditioned to increasing bortezomib doses displayed reduced bortezomib sensitivity and elevated xCT levels. Inhibiting Xc- activity potentiated bortezomib-induced cytotoxicity in myeloma cell lines and primary cells, and re-established sensitivity to bortezomib in bortezomib-conditioned cells. We propose that intracellular GSH level is the main determinant of bortezomib-induced cytotoxicity in a subset of myeloma cells, and that combined targeting of the proteasome and the Xc- cystine-glutamate antiporter can circumvent bortezomib resistance.

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Section: Methodsmentioning

confidence: 99%

Intracellular glutathione determines bortezomib cytotoxicity in multiple myeloma cells

Starheim

Holien

Misund

et al. 2016

Blood Cancer Journal

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“…10 PCR TaqMan assays used were as follows: MYC, Hs00153408_m1; MYCL1, Hs00420495_m1; and GAPDH, Hs99999905_m1 (Applied Biosystems).…”

Section: Cellsmentioning

confidence: 99%

Addiction to c-MYC in multiple myeloma

Holien

Våtsveen

Hella

et al. 2012

Blood

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156

138

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IntroductionMembers of the MYC family are important oncogenes involved in the development of malignant cells. 1 This may also be the case in multiple myeloma (MM), a malignancy of antibodyproducing plasma cells in bone marrow. The activity of c-MYC in MM increases with disease stage. 2,3 The mechanism by which c-MYC is activated in each case is unclear; however, multiple signaling pathways converge on c-MYC. Translocations involving MYC and immunoglobulin genes (IG) are relatively rare in MM and considered late progression events. 4 c-MYC regulates transcription of up to 15% of the genes in human cells by binding to its obligate partner MAX. Many cancer cells may develop a dependency on c-MYC activity; and by preventing this activity, the cells may stop dividing or even undergo apoptosis. In agreement with this, short-hairpin RNA targeting MYC was shown to be lethal to a number of human myeloma cell lines. 5 A small-molecule inhibitor, termed 10058-F4, has been identified that is proposed to specifically inhibit c-MYC-MAX heterodimerization, thereby preventing transactivation of c-MYC target genes. 6,7 The inhibitor has been shown to have growth inhibitory effects on lymphoma and acute myelogenous leukemia cells. 8,9 Methods CellsMyeloma cell lines used in this study were U266, INA-6, JJN-3, KMS-12-BM, IH-1, and KJON. Details on cell culture conditions are found in supplemental Methods (available on the Blood Web site; see the Supplemental Materials link at the top of the online article). CD138 ϩ patient cells obtained through the Norwegian Myeloma Biobank were purified using RoboSep automated cell separator and Human CD138 Positive Selection Kit (Stem Cell Technologies). Bone marrow stromal cells (BMSCs) from patients were obtained by plastic adherence and cultivated as stated in supplemental Methods. The project was approved by the Regional Ethics Committee, and patients gave informed consent in accordance with the Declaration of Helsinki. Patient characteristics are described in supplemental Table 1.Cell viability measurements and quantitative RT-PCR were performed as described previously. 10 PCR TaqMan assays used were as follows: MYC, Hs00153408_m1; MYCL1, Hs00420495_m1; and GAPDH, Hs99999905_m1 (Applied Biosystems).Description of other reagents, immunoblotting, and knockdown experiments is found in supplemental Methods. Results and discussionTo address the c-MYC dependency of myeloma cells, we decided to evaluate the effect of 10058-F4 in myeloma cell lines and primary cells by in vitro studies. First, both mRNA and protein expression of c-MYC and L-MYC was determined in 6 cell lines by quantitative RT-PCR and immunoblotting ( Figure 1A). Five of the cell lines expressed c-MYC, whereas U266 only had L-MYC, as previously reported. 11 L-MYC mRNA was also detected in KMS-12-BM and to a lesser extent in INA-6 and JJN-3, although the levels of L-MYC were negligible compared with c-MYC. The effect on cell viability was evaluated in myeloma cell lines treated with increasing concentrations of 10058-F4 for 48 hour...

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“…To detect chromosomal abnormalities in INA‐6 and IH‐1 cells, metaphase FISH was used, and any IGH rearrangement was detected using fusion strategy. INA‐6 had an unbalanced t(11;14) (19) with two der(14) chromosomes and no der(11) (Fig. S1a).…”

Section: Resultsmentioning

confidence: 99%

“…IH‐1 was used as a positive control in the experiments because of its t(4;14) and unmutated FGFR3 . OH‐2 was used as a negative control because it had no IGH translocations or FGFR3 expression (11). The proliferation assay measuring DNA synthesis showed decreased proliferation of INA‐6 and IH‐1 in the presence of the inhibitor SU5402 at concentrations as low as 3.25 and 15 μ m , respectively (Fig.…”

Section: Resultsmentioning

confidence: 99%

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FGFR3 is expressed and is important for survival in INA‐6, a human myeloma cell line without a t(4;14)

Våtsveen

Brenne

Dai³

et al. 2009

European J of Haematology

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The decreased viability and proliferation in INA-6, following inhibition with FGFR3 inhibitors, indicates that FGFR3 may play a role also in cells without t(4;14) - and hence without high expression of MMSET, the ubiquitous oncoprotein in MM cells with t(4;14). This gives further credibility to the notion that FGFR3 expression is not just an epiphenomenon in t(4;14) MM, but an important part of the malignant phenotype.

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OH-2, a hyperdiploid myeloma cell line without an IGH translocation, has a complex translocation juxtaposing MYC near MAFB and the IGK locus

Cited by 16 publications

References 22 publications

Intracellular glutathione determines bortezomib cytotoxicity in multiple myeloma cells

Intracellular glutathione determines bortezomib cytotoxicity in multiple myeloma cells

Addiction to c-MYC in multiple myeloma

FGFR3 is expressed and is important for survival in INA‐6, a human myeloma cell line without a t(4;14)

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