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Mechanism of CAG Regimen Eliminating T-cell Acute Lymphoblastic Leukemia Cells of A3 Cell Line

Author: WuYanLing
Tutor: SunAiNing;XueShengLi
School: Suzhou University
Course: Internal Medicine
Keywords: Leukemia T cells Acute CAG regimen A3 cell lines Signal transduction Ras-MAPK PD98059
CLC: R733.7
Type: Master's thesis
Year: 2008
Downloads: 14
Quote: 0
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Purpose (1) research the WPW program (CAG program) Clear T-cell acute lymphoblastic leukemia (T-ALL) cell lines A3 cell mechanism and granulocyte colony-stimulating factor and its receptor (G-CSF/G-CSFR ) the role played by the ligand receptor system in this mechanism. The method (1) with the G-CSFR monoclonal antibody labeled binding flow cytometry A3 cell surface expression levels of G-CSFR. (2) A3 cells as an in vitro experimental model, the packets were added to different concentrations of G-CSF (5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml; 0ng/ml as control group), incubated for 48 incubator hours later, the cells were collected, propidium iodide nuclear staining, flow cytometry the A3 cell cycle changes. (3) the role of different concentrations of cytarabine (Ara-C) combined with G-CSF A3 cells after 48 hours using Cell Counting Kit (CCK-8) kit detects drugs A3 cell inhibition rate. (4) AnnexinV kit combines flow cytometry Ara-C, the A3 changes in the level of apoptosis Aclacinomycin (ACR) and the combined effects of G-CSF after 48 hours. (5) MEK in the Ras-MAPK pathway specific inhibitor PD98059 and G-CSF total role A3 cells after 48 hours by flow cytometry analysis of cell cycle changes of CCK-8 cell viability. Results (1) flow cytometry measured A3 cell surface G-CSFR expression rate of about 94.2%, SPSS 13.0 statistical analysis of KG-1 cells and the positive control group, acute myeloid leukemia cell strains (AML), no significant difference. (2) the proportion of cells in a certain concentration range of G-CSF after 48h S of A3 is gradually increased with the increase of the concentration of G-CSF and the G-CSF concentration of 15ng/ml highest, and the difference with the control group with statistical significance, and then with the G-CSF is increased but decreased gradually. (3) The role of the drug group after 48h, CCK-8 measured Ara-C G-CSF group than in the single-agent Ara-C group more pronounced inhibition of the growth of A3 cells (Ara-C concentration of 10-5M and 10-6M P lt ; 0.05). (4) AnnexinV apoptosis detection kit the A3 early apoptosis percentage results: Ara-C ACR the G-CSF drug group cell early apoptosis rate of about 38%, significantly higher than Ara-C ACR drug group, the difference has statistical significance. (5) With the gradually increasing concentration PD98059 A3 cells in S-phase fraction and A3 cells OD value decreased, respectively PD98059 concentration from 40μM ~ 100μM and 50μM ~ 100μM significant difference compared with the control group (p lt; 0.05). Conclusion (1) A3 cell surface of T-ALL cell lines may be highly expressed in the G-CSFR. (2) G-CSF/G-CSFR ligand receptor system in the process of CAG regimen cleared A3 cells, synergy with chemotherapy drugs, by mobilizing the G0 phase cells enter the cell proliferation cycle, enhance the sensitivity of chemotherapy drugs to cells thus cleared Cell. (3) induction of apoptosis may remove one of the mechanisms of T-ALL cell lines A3 cells CAG regimen. (4) binding of G-CSF and Effect of specific cell surface receptor (G-CSFR), by activating a series of downstream signal transduction pathways, play a role in the Ras-MAPK pathway is one; PD98059 possible through specific inhibition of the Ras -MAPK pathway MAPK phosphorylation in to prevent downstream signal transduction thereby partially inhibit the effects of G-CSF.

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CLC: > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Leukemia
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