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Study on the Mechanisms of Leukemia Cells’ Resistance to Chemotherapeutic and Biotherapeutic Agents Mediated by Glutathione S-transferase

Author: LinRongRong
Tutor: ChenHanChun;ZhangDianZheng
School: Central South University
Course: Biochemistry and Molecular Biology
Keywords: Chronic myeloid leukemia Interferon Hydroxyurea GSTPi
CLC: R733.72
Type: Master's thesis
Year: 2010
Downloads: 16
Quote: 0
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Abstract


Chronic myeloid leukemia (chronic myelogenous leukemia, CML) is a class of hematological malignancies. Hydroxyurea; interferon-a (interferon-alpha, IFN-α) (hydroxyurea, HU), the CML preferred one of the chemotherapy drugs is effective biological therapeutic agent of hematopoietic malignancies and lymphomas; CML cells both presents resistance. Glutathione S-transferase (glutathione S-transferases Pi, GSTPi) is a member of the phase II drug metabolizing enzymes, may be involved in the formation process of CML cells resistant to chemotherapy and / or biological therapeutic agent. Objective: This subject is based on the CML cell line (K562, KT-1/A3 KT-1/A3R) cells as an experimental model to study the HU, IFN-α, or both combined CML cell proliferation, apoptosis, analysis HU, IFN-α alone or combined effects before, the CML cells GSTPi after expression differences, to explore GSTPi role in the formation process of CML cells resistant to chemotherapy and / or biological therapeutic agent and its possible mechanism. Method: 1. Cell count analysis of K562, KT-1/A3 and KT-1/A3R cell control group and drug group cell growth state, cell growth curve; 2.MTT assay of K562, KT-1/A3 and KT-1/A3R cells in the control group and drug effects growth inhibition rate after 48 hours; flow cytometry K562 KT-1/A3 and KT-1/A3R cell cycle and apoptosis rate; 4. RT -PCR and Western-blotting detection K562, the expression level of the control group and the role of the drug in KT-1/A3, and KT-1/A3R cells after 48 hours of GSTPi. In Results: 1.HU to inhibit the K562, KT-1/A3, and KT-1/A3R cell proliferation and promote the apoptosis compared with the control group were statistically significant (P lt; 0.01); 2.HU joint IFN-a synergistic inhibition of K562, KT-1/A3, and KT-1/A3R cell proliferation and induce apoptosis in CML cells, compared with monotherapy, combination therapy better inhibition of cell proliferation, promote apoptosis (P lt RT-PCR results show that, in the HU under the K562, KT-1/A3R cells compared with the control group, GSTPi gene expression is significantly reduced (P lt; 0.05);; 0.05); 4 and Western blotting results show that, the HU can cut of K562, KT-1/A3R cells of GSTPi protein levels (P lt; 0.05). Conclusion: 1.HU, and HU United IFN-α could significantly inhibit the proliferation of CML cells, and induction of apoptosis; HU and IFN-α than either used alone inhibitory effect on the proliferation of CML cells and induce apoptosis in CML cells effect more obvious; GSTPi the expression level related to the K562, KT-1/A3R interferon resistance, HU down GSTPi expression levels; 3.K562, KT-1/A3R cells to IFN-α-resistant, and HU or HU in combination with IFN-α can be alleviated the K562, KT-1/A3R cells resistant to IFN-α, and combination therapy can synergistically inhibit K562, KT-1/A3R cell proliferation and induce apoptosis.

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