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Multidrug resistance reversal agents screening and shikonin anti- the MDR tumor activity of research

Author: FangJianPing
Tutor: HuXun
School: Zhejiang University
Course: Biochemical Engineering
Keywords: Multidrug resistance MDR Drug screening Shikonin Apoptosis Anti-tumor multidrug resistance
CLC: TQ461
Type: Master's thesis
Year: 2004
Downloads: 206
Quote: 1
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Abstract


Tumor multidrug resistance (Multidrug Resistance, MDR) has become a chemotherapy obstacles, is also plagued by a major problem for oncologists and cancer patients. The mechanism of MDR transporter protein PgP intracellular drug accumulation in tumor cells caused by overexpression of MRP, LRP and MXR reduction, enhanced intracellular detoxification system capacity, as well as changes in drug targets. One of the most typical is due to overexpression of ATP-dependent efflux \MDR the human erythroleukemia K562/ADR cell model in this paper in order to establish the PgP overexpression of 30 kinds of Chinese herbal monomer sample library established directional filtering using the MTT assay method, in order to obtain a reversal of MDR activity monomer and further study of its pharmacological effects. 1. Anticancer drugs the doxorubicin induced the K562 establish MD subline K562/ADR, has been tested to 7.8 times the resistance to adriamycin, vincristine resistance more than 60 times, epirubicin 98 times. And after the detection of the resistance spectrum, the accumulation of intracellular doxorubicin and PGP detection of three means are also shows, the MDR mechanism of the cell model is a result of the excessive expression of membrane proteins of Pgp, its expression, and negative control group compared to as high as 88.79%. 2. MDR reversing agents, screening and rescreening the MDR cell model K562/ADR sample libraries by the 30 kinds of Chinese herbal medicine monomer, we found that andrographolide 6μg/ml concentration no cytotoxicity (survival> 90%) , a certain role in the reversal of MDR, but the effect is not strong. Tanshinone Ⅰ, cryptotanshinone of dihydro tanshinones I shikonin cell killing effect, while the strongest killing effect of shikonin honokiol both destruction and reversal of role, but not very strong. 3. The determine Shikonin as the research object, we found that the the Boraginaceae perennial herb comfrey active ingredients monomer Shikonin has very strong anti-multidrug resistant tumor cells. Cells induced by shikonin, Ladder and flow cytometry analysis Figure Central Asian diploid peak emergence of genomic DNA agarose gel electrophoresis and stained oil immersion observed changes in cell morphology, and Caspase-3 activation of the enzyme, and these show that the shikonin the antitumor mechanism induced apoptosis, and by comparison has the characteristics the parental cells K562 and the resistant cells K562/ADR effect quite. 50 by killing effect obtained IC of the MTT assay Shikonin two cells very close, were 0.436μg/ml and 0.445μg/ml HPLC detection of intracellular drug accumulation and retention concentration show that The shikonin be absorbed into a certain compound, and the concentration of the compound over time change the relationship of Zhejiang University, a master's degree thesis on the two types of cells is consistent. This further illustrates shikonin quite two cells. Conclusion: drug screening, we found that a Chinese herbal monomer shikonin induced sensitive K562 and MDR cell line K562 from the anti-tumor effect of DR apoptosis, and the effects of the two quite overcome MDR characteristics. And the possible induction of apoptotic pathways are discussed and put forward the idea of ??the next step, some not yet clear conclusion.

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