Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0026764 (multiple myeloma)
 36,148 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Prior work has reported that cotransfecting a gene of interest with the selectable marker neo can seriously perturb a number of cellular processes. In this study the influence of the neo gene on the growth, death, and metabolism of a murine myeloma NS0 cell line, expressing a chimeric antibody, was investigated. A pool of neo transfectants, 6A1-NEO, was selected with 500 microg/mL G418. Quantitative PCR analysis revealed that 6A1-NEO contained, on average, three copies of the neo gene per cell. Batch cultivation of 6A1-NEO showed that there was a 36% increase in maximum viable cell concentration, a 20% increase in the maximum apparent growth rate, and a 134% increase in cumulative cell hours as compared with the parent, 6A1-(100)3. Batch cultivation of five randomly selected clones illustrated that 6A1-NEO's advantage over the parent was not due to clonal variation. Neither the use of G418 during the selection process nor the cultivation of cells in the presence of G418 were responsible. This implied that the neo gene product, APH(3')-II, was causing the changes in proliferative capacity. Analysis of the cell cycle revealed that there were no differences in the distribution of cells in the G(1), S, and G(2) phases. When cell growth was synchronized, there were no observed differences in cell-cycle duration. 6A1-NEO resisted the onset of apoptosis during the growth phase. Consequently, there was a larger viable population of 6A1-NEO cells available for proliferation as compared with the parent. However, 6A1-NEO died at the same rate as the parent when resuspended in spent media or after treatment with staurosporin. Expression of the anti-apoptotic protein Bcl-2 was upregulated in 6A1-NEO, indicating that APH(3')-II could be acting by modulating endogenous gene expression. Analysis of key metabolites showed that 6A1-NEO's specific glucose consumption rate was 133% higher, whilst its specific glutamate consumption rate was 45% lower than the parent. 6A1-NEO's efficient utilization of glutamate and shift towards glucose metabolism may have contributed to the rise in proliferative capacity. However, this was accompanied by a 70% drop in the specific antibody production rate. These results show that the increase in growth rate and proliferative capacity caused by the expression of recombinant APH(3')-II was associated with changes in metabolism, apoptosis, and endogenous gene expression.
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PMID:Enhanced growth in NS0 cells expressing aminoglycoside phosphotransferase is associated with changes in metabolism, productivity, and apoptosis. 1626 48

This study was aimed to investigate the effect of VEGF antisense RNA on proliferation and apoptosis in myeloma cell line U266 as well as on angiogenesis in endothelial cell ECV304, and to explore the feasibility of gene therapy for multiple myeloma using VEGF antisense RNA. The VEGF121 cDNA was inserted into multiple clone site of eukaryotic expression vector pIRES2-EGFP to construct the recombinant plasmid AS-VEGF. Restriction endonuclease analysis and DNA sequencing were used to confirm the reverse orientation of the VEGF cDNA. The recombinant plasmid was transfected into human myeloma cell line U266 and the positive clone was screened by G418. The VEGF mRNA and protein expressions of the positive clone were detected by RT-PCR and Western blot respectively. The viability and apoptosis of U266 cells were observed by MTT assay, flow cytometry. Angiogenesis was tested by network formation of endothelial cells on matrigel. The results indicated that the recombinant plasmid AS-VEGF expressing VEGF antisense RNA were constructed successfully. VEGF expression in U266 cells was blocked partially by VEGF antisense RNA. Expression of VEGF mRNA and protein decreased more significantly in U266 cells transfected by AS-VEGF than that in control group. Then increasing of apoptosis and inhibition of proliferation in U266 cells transfected by AS-VEGF were observed. Vasoformation on matrigel in the supernatants of U266 culture group transfected by AS-VEGF decreased more significantly than that in control group. It is concluded that VEGF antisense RNA can inhibit the expression of VEGF gene in U266 cells, thereby inhibits the proliferation of U266 cells; increases the apoptosis of U266 cells; and inhibits angiogenesis in vitro.
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PMID:[Effect of VEGF antisense RNA on inducing apoptosis of myeloma cells and inhibition of angiogenesis in endothelial cells in vitro]. 1842 55


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