Establishment of five human myeloma cell lines

Namba, Masayoshi; Ohtsuki, Tatsuyuki; Mori, Masaharu; Togawa, Atsushi; Wada, Hideho; Sugihara, Takashi; Yawata, Yoshihito; Kimoto, Tetsuo

doi:10.1007/bf02623725

Cited by 72 publications

(51 citation statements)

References 31 publications

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“…The KMS-11, KMS-12 and KMS-18 cell lines have been previously described (Namba et al, 1989;Otsuki et al, 1998); the NCI-H929 and OPM-2 cell lines were obtained from DSMZ (Braunschweig, Germany); the U266 cell line was obtained from the American Type Culture Collection (Rockville, MD, USA). The cell lines were cultured at 378C in a humidi®ed atmosphere containing 5% CO 2 , and maintained in Iscove's Modi®ed Dulbecco's Medium (IMDM) supplemented with 10% fetal calf serum (FCS).…”

Section: Cell Linesmentioning

confidence: 99%

Deregulated FGFR3 mutants in multiple myeloma cell lines with t(4;14): comparative analysis of Y373C, K650E and the novel G384D mutations

et al. 2001

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The t(4;14)(p16.3;q32) chromosomal translocation occurs in approximately 20% of multiple myelomas (MM) and leads to the apparent deregulation of two genes located on 4p16.3: the ®broblast growth factor receptor 3 (FGFR3) and the putative transcription factor WHSC1/ MMSET. Interestingly, FGFR3 mutations known to be associated with autosomal dominant human skeletal disorders have also been found in some MM cell lines with t(4;14) but their pathogenetic role in MM is still controversial. Since cell lines may represent useful models for investigating the e ects of deregulated FGFR3 mutants in MM, we analysed the expression, activation, signaling pathways and oncogenic potential of three mutants identi®ed so far: the Y373C and K650E in the KMS-11 and OPM-2 cell lines respectively, and the novel G384D mutation here identi®ed in the KMS-18 cell line. All of the cell lines present a heterozygous FGFR3 gene mutation and transcribe the mutated allele; unlike KMS-11 and OPM-2 (which express the IIIc isoform), the KMS-18 cell line expresses prevalently the isoform IIIb. We demonstrated that, under serum-starved conditions, KMS-11 and OPM-2 cells express appreciable levels of phosphorylated FGFR3 mutants indicating a constitutive activation of the Y373C and K650E receptors; the addition of the aFGF ligand further increased the level of receptor phosphorylation. Conversely, the FGFR3 mutant in KMS-18 does not seem to be constitutively activated since it was phosphorylated only in the presence of the ligand. In all three MM cell lines, ligand-stimulated FGFR3 mutants activated the MAP kinase signaling pathway but did not apparently involve either the STAT1 or STAT3 cascades. However, when transfected in 293T cells, G384D, like Y373C and K650E, was capable of activating MAPK, STAT1 and STAT3 under serum-starved condition. Finally, a focus formation assay of NIH3T3 cells transfected with FGFR3-expressing plasmid vectors showed that Y373C and K650E (albeit at di erent levels) but not G384D or the wild-type receptor, can induce transformed foci. Overall, our results support the idea that FGFR3 mutations are graded in terms of their activation capability, thus suggesting that they may play a critical role in the tumor progression of MM patients with t(4;14). Oncogene (2001) 20, 3553 ± 3562.

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Section: Cell Linesmentioning

confidence: 99%

Deregulated FGFR3 mutants in multiple myeloma cell lines with t(4;14): comparative analysis of Y373C, K650E and the novel G384D mutations

et al. 2001

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“…14,15 The Granta line was derived from a human mantle cell lymphoma and was a kind gift from DM Jadayel, 16 Institute of Cancer Research, Surrey, UK. Cells were grown at 37°C (5% CO 2 ) in RPMI 1640 media supplemented with 10% (v/v) heat-inactivated fetal bovine serum, 2 mM/l L-glutamine, and 100 U/ml penicillin and 100 g/ml streptomycin.…”

Section: Cell Culturementioning

confidence: 99%

Frequent inactivation of the cyclin-dependent kinase inhibitor p18 by homozygous deletion in multiple myeloma cell lines: ectopic p18 expression inhibits growth and induces apoptosis

et al. 2002

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Multiple myeloma (MM) is a clonal neoplasm of plasma cells which offers an excellent model to study multistep molecular oncogenesis. In 20-25% of primary tumors and cell lines examined, cyclin D1 is overexpressed due to the translocation t(11;14)(q13;q32). We have characterized cyclin-dependent kinase inhibitor p15 (CDKN2B), p16 (CDKN2A) and p18 (CDKN2C) deletions in cyclin D1-expressing and non-expressing MM cell lines. p18 was found to be frequently deleted (38%); in some cases p18 deletions coexisted with hemizygous p16 deletion. To examine the function of p18 as a putative tumor suppressor in myeloma cells, a zinc-inducible p18 construct was stably transfected into KMS12, a MM cell line with biallelic p18 and monoallelic p16 deletions as well as cyclin D1 overexpression. Ectopic expression of p18 caused 40-45% growth suppression as determined by trypan blue exclusion and MTS assays. p18 induction also resulted in apoptosis, suggesting that inhibition of the cyclin D1/CDK/pRb pathway in these tumor cells could be a crucial step toward the induction of tumor regression via apoptotic cell death. This cell cycle pathway is thus frequently mutated and provides a potentially novel target for gene therapeutic or pharmacologic approaches to human myeloma.

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“…46 In the present study, we used five human myeloma cell lines including KMS-18, established at Kawasaki Medical School, [47][48][49] and one of well characterized myeloma cell line, U266, 50 treated with mycoplasma removal agent (MRA) 51,52 to observe ammonia production in vitro. These cell lines produce and release significantly higher amounts of ammonia into culture medium (13.65 ± 5.84 mol/l/10 × 10 3 proliferating cells) than non-myeloma hematological cell lines, or the HepG2 human hepatic carcinoma cell line (4.16 ± 1.90 mol/l/10 × 10 3 proliferating cells).…”

Section: Introductionmentioning

confidence: 99%

In vitro excess ammonia production in human myeloma cell lines

et al. 1998

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It is well known that cases with multiple myeloma reveal various clinical manifestations such as pancytopenia, hyperproteinemia, renal dysfunction, bone lesions, hypercalcemia and immunodeficiency. Recently, a few more clinical features associated with myeloma, such as salivary type hyperamylasemia and elevated serum C-reactive protein (CRP) concentration, have been reported. The elevation of CRP is thought to be related to interleukin-6 (IL-6) production by myeloma cells, because of identification of IL-6 as an autocrine and/or paracrine growth factor for myeloma cells. More recently, there have been several reports of cases with myeloma associated with hyperammonemia. This hyperammonemia is not considered to be due to liver dysfunction, because in most of these cases tests revealed normal hepatic function, and some cases showed different patterns of serum amino acid distribution than that associated with hepatic failure. However, there have been no apparent observations of ammonia production by myeloma cells. In this study, we used six human myeloma cell lines including KMS-18, which was recently established from a myeloma case associated with hyperammonemia. These lines were treated with MRA (mycoplasma removal agent) to observe ammonia production in vitro. They produced and released significantly higher levels of ammonia into culture medium than non-myeloma hematological cell lines or the HepG2 human hepatic carcinoma cell line. Although attempts to analyze the relative expression levels of the enzymes related to ammonia biosynthesis using the reverse transcriptase-polymerase chain reaction assay failed to detect any differences between these myeloma lines and other cell lines, in vitro excess ammonia production by the myeloma cells was confirmed and the relevance to clinical manifestations is discussed.

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Establishment of five human myeloma cell lines

Cited by 72 publications

References 31 publications

Deregulated FGFR3 mutants in multiple myeloma cell lines with t(4;14): comparative analysis of Y373C, K650E and the novel G384D mutations

Deregulated FGFR3 mutants in multiple myeloma cell lines with t(4;14): comparative analysis of Y373C, K650E and the novel G384D mutations

Frequent inactivation of the cyclin-dependent kinase inhibitor p18 by homozygous deletion in multiple myeloma cell lines: ectopic p18 expression inhibits growth and induces apoptosis

In vitro excess ammonia production in human myeloma cell lines

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