Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT01586 (G418)
 2,237 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease of transformed hematopoietic progenitor cells. It is now clear that the chimeric bcr/abl P210(bcr/abl) fusion protein, which is generated by the reciprocal translocation t (9; 22), inhibits apoptosis and increase proliferation. P210(bcr/abl) plays a central role in the pathophysiology of CML. The purpose of this study was to construct a cell line model that bcr/abl expression can be regulated by Tet-off inducing-expression-system. The full-length b3a2 bcr/abl cDNA was subcloned into the pTRE2hyg expression vector to construct the pT2-P210 plasmid. 293 cells were firstly transfected with Tet-off plasmid and the clone that the Tet-off system can work effectively after transfected with pTRE2hyg-LUC was selected by luciferase activity assay. The pT2-P210 plasmid was then transfected into the selected clone and cells were then selected for hygromycin B and G418 resistance. The results showed that individual subclones expressing bcr/abl after withdrawing doxycycline were 293pT2-P210 cell line. In conclusion, selected 293pT2-P210 cells are cells that bcr/abl expression can be regulated by Tet-off inducing-expression-system. They are suitable thoroughly to study the function of bcr/abl fusion gene and its signal regulation mechanism.
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PMID:[Construction of 293pT2-P210 cell line enables expression of bcr/abl to be regulated by Tet-off inducing-expression-system]. 1749 20

This study was purposed to explore the effect of specific small interfering RNA targeting at bcr-abl fusion gene and its combination with p27 gene clone on the proliferation, cell cycle and apoptosis of chronic myeloid leukemia (CML) cell line K562. CML cell line K562 was used as the study object. A 21nt siRNA targeting at the fusion site of b3:a2 mRNA in bcr-abl fusion gene was designed, synthesized and transfected into the K562 cells as RNA interference group. Northern blot was used to detect the bcr-abl fusion gene, Western blot was used to detect the expression of P210 protein and apoptosis-related protein BCL-xL after the transfection. Meanwhile, p27 gene was amplified from peripheral blood mononuclear cells by RT-PCR, and was confirmed to be correct by sequencing, then p27-pcDNA3.1 vector was constructed and transfected into K562 cell line by Lipofectin. After being selected with G418, p27-pcDNA3.1-K562 cell clone stably expressing p27 was isolated. P27 protein was identified by Western blot. Finally, siRNA and p27 gene clone were together applied to K562 cells, the cell survival rate was tested by MTT. The cell cycle and the apoptosis were tested by flow cytometry. The result showed that in contrast with the control group, the expression level of bcr-abl fusion gene was much lower in siRNA group, about 18.4% of K562 cells in siRNA group were apoptotic at 24 hours after siRNA transfection, and the expression of apoptosis-associated protein BCL-xL was greatly down-regulated. The expression of P27 protein could be detected by Western blot in p27-pcDNA3.1-K562 cells. The strong inhibition of cell proliferation was observed in p27-pcDNA3.1-K562 cells, as compared with control K562 cells. The count of p27-pcDNA3.1-K562 cells in G(0)/G(1) phase increased apparently, but that in S phase declined greatly. Cell cycle was arrested in G(0)/G(1) phase. After the combination of p27-pcDNA3.1-K562 cells with specific siRNA, the percentage of apoptosis obviously increased and cell survival rate significantly declined. It is concluded that the specific siRNA distinctly inhibits the expression of bcr-abl fusion gene, and can induce K562 cell apoptosis. The combination of specific siRNA with P27 gene clone displays a synergy of inhibition and pro-apoptosis effects to K562 cells.
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PMID:[Effects of specific siRNA targeting at bcr-abl fusion gene and its combination with p27 gene clone on chronic myeloid leukemia cell line K562]. 2056 11