Genome-Wide Analysis of Mouse Myeloma Cell Lines Expressing Therapeutic Antibodies

2007 ◽  
Vol 23 (4) ◽  
pp. 911-920 ◽  
Author(s):  
Haimanti Dorai ◽  
Katherine Li ◽  
C. Chris Huang ◽  
Anton Bittner ◽  
Jose Galindo ◽  
...  
Keyword(s):  
Cell Lines ◽  
Myeloma Cell ◽  
Therapeutic Antibodies ◽  
Genome Wide Analysis ◽  
Mouse Myeloma Cell ◽  
Genome Wide ◽  
Mouse Myeloma

scholarly journals Differential Extraction of DNA Polymerase from the Nucleus of the Mouse Myeloma Cell

1979 ◽  
Vol 4 (4) ◽  
pp. 295-306 ◽  
Author(s):  
Akio Matsukage ◽  
Nobuyuki Nishioka ◽  
Miwako Nishizawa ◽  
Taijo Takahashi
Keyword(s):  
Dna Polymerase ◽  
Myeloma Cell ◽  
Differential Extraction ◽  
Mouse Myeloma Cell ◽  
Mouse Myeloma

Nucleotide sequence of an unequal sister chromatid exchange site in a mouse myeloma cell line

1989 ◽  
Vol 9 (3) ◽  
pp. 1324-1326
Author(s):  
D R Katzenberg ◽  
S A Tilley ◽  
B K Birshtein
Keyword(s):  
Nucleotide Sequence ◽  
Cell Line ◽  
Sister Chromatid ◽  
Sister Chromatid Exchange ◽  
Present Report ◽  
Myeloma Cell ◽  
Myeloma Cell Line ◽  
Mouse Myeloma Cell ◽  
Mouse Myeloma Cell Line ◽  
Mouse Myeloma

The mouse myeloma cell line MPC 11 carries two C gamma 2a immunoglobulin heavy-chain genes on the expressed chromosome, a duplication shown to have occurred through unequal sister chromatid exchange (USCE). In the present report, we present the nucleotide sequence of the USCE joint and show that both breaks occurred within tracts of repeated TC dinucleotides. Additional TC dinucleotide tracts and two oligonucleotide segments (N sequences) were inserted at the USCE site.

Biochemical characterization of the cholesterol-dependent growth of the NS-1 mouse myeloma cell line

1986 ◽  
Vol 163 (1) ◽  
pp. 117-126 ◽  
Author(s):  
Jan-Kan Chen ◽  
Tetsuji Okamoto ◽  
J.Denry Sato ◽  
Gordon H. Sato ◽  
Don B. McClure
Keyword(s):  
Cell Line ◽  
Biochemical Characterization ◽  
Myeloma Cell ◽  
Myeloma Cell Line ◽  
Mouse Myeloma Cell ◽  
Mouse Myeloma Cell Line ◽  
Dependent Growth ◽  
Mouse Myeloma

scholarly journals Mouse myeloma cells that make short immunoglobulin heavy chains: pleiotropic effects on glycosylation and chain assembly.

1984 ◽  
Vol 98 (6) ◽  
pp. 2215-2221 ◽  
Author(s):  
A L Kenter ◽  
T Warren ◽  
D Shields ◽  
B K Birshtein
Keyword(s):  
Conformational Changes ◽  
Myeloma Cell ◽  
In Vitro Translation ◽  
Myeloma Cell Line ◽  
Present Evidence ◽  
Heavy Chains ◽  
Mouse Myeloma Cell ◽  
Mouse Myeloma Cell Line ◽  
Mouse Myeloma

Two variants in immunoglobulin heavy chain production, derived from the MPC 11 mouse myeloma cell line, make short heavy (H) chains with identical precise deletions of the CH3 domain. The CH3 domain is expressed in the H chain mRNA from both variants. Although in vitro translation of this mRNA produces one H chain species, deleted heavy chains are secreted as heavy-light (HL) and H2L2 moieties in contrast to MPC 11, which secretes only H2L2 . The heavy chains of HL apparently contain more carbohydrate (CHO+) than do the H chains of H2L2 , and inhibition of N-linked glycosylation results in the secretion of relatively more H2L2 . Here we present evidence suggesting that (a) the absence of the CH3 domain has led to conformational changes in these molecules, (b) these changes permit posttranslational glycosylation, and (c) unrestrained glycosylation can frequently yield unusual CHO+ structures that make complete assembly unlikely.

A soluble biotinyl-α-amanitin affinity probe for the isolation of RNA polymerase B

1986 ◽  
Vol 64 (9) ◽  
pp. 923-929 ◽  
Author(s):  
A. C. Vaisius ◽  
H. Faulstich
Keyword(s):  
Cell Culture ◽  
Rna Polymerase ◽  
Potent Inhibitor ◽  
Mammalian Cell Culture ◽  
Sodium Dodecyl ◽  
Myeloma Cell ◽  
Affinity Probe ◽  
Gel Beads ◽  
Mouse Myeloma Cell ◽  
Mouse Myeloma

Utilizing the 6′-hydroxyindole moiety of α-amanitin for substitution, biotinyl-α-amanitin has been synthesized to use as a soluble affinity probe for the isolation of RNA polymerase B from mammalian cell culture. The synthetic biotinyl-α-amanitin remains a potent inhibitor of RNA polymerase B having a Ki of 4.1 × 10−8 M as compared with a Ki of 5 × 10−9 M for natural α-amanitin. RNA polymerase B complexed with biotinyl-α-amanitin can be isolated on Bio-Gel P300 polyacrylamide gel beads to which avidin has been attached. RNA polymerase B may then be released from the complex by treatment with sodium dodecyl sulfate or by monochromatic irradiation at 314 nm which destroys the amatoxin moiety. We have used this affinity probe to analyze the subunit composition of RNA polymerase B from various mouse myeloma cell lines. We believe that the biotinyl-α-amanitin may be very useful for the isolation of factors which associate with RNA polymerase B; e.g., we have substantiated that actin can be associated with RNA polymerase.

Biological and Molecular Characterization by Integrative Genomics Approach of CMA-03/06, a Newly Established Interleukin-6 Independent Variant of the CMA-03 Human Myeloma Cell Line.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1814-1814
Author(s):  
Donata Verdelli ◽  
Lucia Nobili ◽  
Katia Todoerti ◽  
Laura Mosca ◽  
Sonia Fabris ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Line ◽  
Molecular Characterization ◽  
Cell Lines ◽  
Stromal Cells ◽  
Interleukin 6 ◽  
Myeloma Cell ◽  
Growth And Survival ◽  
Genome Wide

Abstract Abstract 1814 Poster Board I-840 Background The growth and survival of multiple myeloma (MM) cells in the bone marrow microenvironment is regulated by functional complex interactions between the tumor cells and the surrounding bone marrow stromal cells mediated by adhesion molecules and the production of several cytokines of which interleukin-6 (IL-6) has been identified as the most important. Major advances in the investigation of MM biology were made possible by the availability of human myeloma cell lines (HMCLs). The IL-6-dependent CMA-03 cell line was established in our laboratory from a peritoneal effusion of a refractory relapsed MM patient. By gradually decreasing the IL-6 added to the culture, an IL-6-independent variant, CMA-03/06, could be obtained. Aims. To perform a biological and molecular characterization of this novel cell line, and to provide insights into the signaling pathways and target genes involved in the growth and survival of CMA-03/06. Methods. The growth, immunophenotypic, cytogenetic and fluorescence in situ hybridization (FISH) characterization of CMA-03/06 cell line was performed by means of standard procedures. IL-6 production into the culture media was determined using a high sensitivity IL-6 specific ELISA. Genome-wide profiling data were generated by means of Affymetrix GeneChip® Human Mapping 250K Nsp arrays; copy number (CN) alterations were calculated using the DNAcopy Bioconductor package, based on circular binary segmentation method. Global gene expression profiling (GEP) was performed by means of the GeneChip® Human Gene 1.0 ST Arrays (Affymetrix); the supervised analyses were done using the SAM software version 3.0. Results Unlike CMA-03, the addition of IL-6 to the culture medium of CMA-03/06 cells or co-culture with multipotent mesenchymal stromal cells did not induce an increase in CMA-03/06 proliferation. IL-6 was not detected in the supernatants from either CMA-03 or CMA-03/06 cell lines within 48 h, suggesting that the IL-6 independence of CMA03/06 cells is not a result of the development of an autocrine IL-6 loop. Nevertheless, IL-6 induced the activation of STAT3 and STAT1 in both cell lines, even if a slight constitutive STAT3 phosphorylation was found in CMA-03/06. The immunophenotypic analysis showed a significant difference in the expression of three antigens in the 2 cell lines: CD45 was considerably reduced in CMA-03/06 cells, whereas they were found positive for both chains of IL-6 receptor, CD126 and CD130, almost undetectable in CMA-03. Conventional cytogenetic and FISH analyses did not reveal differences between the 2 HMCLs. The genome-wide analysis allowed the identification of about 100 altered chromosomal regions common to both HMCLs, mostly DNA gains. Comparison of CMA-03/06 and CMA-03 cells evidenced a different CN in only 15 small chromosomal regions, 8 of which did not contain any transcript, whereas few genes were located on the other ones. GEP analysis of CMA-03/06 compared with CMA-03 identified 21 upregulated and 47 downregulated genes, many of which particularly relevant for MM biology, mainly involved in cellular signaling, cell cycle, cell adhesion, cell development, regulation of transcription, immunologic, inflammatory or defense activity, apoptosis. None of the genes differentially expressed in CMA-03/06 compared with CMA-03 except 1 were positioned on the chromosomal regions showing a different CN. Finally, CMA-03/06 cell line showed a lower susceptibility to camptothecin-induced apoptosis compared to CMA-03 cells. Conclusions Our data show the IL-6 independence of CMA-03/06 cell line in the absence of an autocrine IL-6 loop; the cells, however, maintain the IL-6 signaling pathway responsiveness. A consistent number of genes particularly relevant for MM biology were found deregulated in CMA-03/06 cell line compared with CMA-03. Furthermore, CMA-03/06 cell line shows an increased resistance to apoptosis. The novel CMA03/06 cell line may thus represent a suitable model for studies investigating molecular mechanisms involved in clonal evolution towards IL-6 and/or stroma-independent growth and survival of myeloma cells. Disclosures No relevant conflicts of interest to declare.

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