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The Expression of MDR1 and BCRP in Breast Cancer Cell Line MCF-7 after Chemotherapeutical Drug Intervention and Its Value in Clinical Practice

Author: MingJie
Tutor: HuangZuo
School: Huazhong University of Science and Technology
Course: Surgery
Keywords: Multidrug resistance MCF-7 cell line Chemotherapy drugs Multidrug resistance gene MDR-1 Breast cancer resistance gene BCRP
CLC: R737.9
Type: Master's thesis
Year: 2007
Downloads: 70
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Abstract


Objective: In vitro evaluation of anthracycline and taxane chemotherapy drugs and breast cancer resistance gene MDR1 and BCRP expression. Method: 1, in vitro cultured human breast cancer MCF-7 cells and inoculated into sterile cell culture plates, treated with various concentrations of doxorubicin, epirubicin, paclitaxel and docetaxel, respectively, to intervene in the MCF-7 cells at different times MDR1, BCRP gene expression levels detected by RT-PCR and PCR product electrophoresis analysis method. 2, real-time quantitative PCR technique further detection of MDR1, BCRP gene expression in different concentrations of doxorubicin and epirubicin different time periods after the intervention, and evaluation of the amount of concentration of anthracycline intervention and intervention time and gene expression effect relationship. 3, The Affiliated Union Hospital, Huazhong University of Science and Technology of breast thyroid surgery in the period from January 2004 to December 2005 to accept the \highly sensitive screening of anthracyclines and anthracycline-resistant breast cancer tissue samples, detection including MDR1, BCRP gene expression differences in clinical significance. 4, the MCF-7, the proportion of stem cells by flow cytometry after the intervention of the different concentrations of doxorubicin and were detected MDR1 gene expression changes in subcellular group, to explore the possible mechanism in the formation of multi-drug resistant . Results: the anthracycline drug intervention and breast cancer cells MCF-7 MDR1 gene expression levels between closely related, MDR1 gene along with the increase of the concentration of the anthracycline and the role of time extension also increases its expression. Anthracycline BCRP gene and taxane and of MDR1, of BCRP are no obvious dose-effect relationship. Anthracyclines after the intervention of breast cancer cells MCF-7 BCRP gene expression law, after the intervention of the taxane MCF-7 MDR1 and BCRP gene expression levels also no significant difference. In addition, in contrast to the MCF-7 cells after the intervention of two anthracycline MDR1 and BCRP gene expression, found no significant difference between the two drugs. And highly sensitive tumor anthracycline MDR1 expression in vitro susceptibility to be significantly lower than the of anthracycline resistant tumor. In addition, control flow cytometric sorting the results and the corresponding cell subsets MDR1 gene level, found that with the increase in the concentration of doxorubicin drug intervention and prolonged duration of action in MCF-7 cells, the stem cell pool enrichment, at the same time, CD44 ~ CD24 ~ - and CD44 to CD24 cell populations ~~ two subgroups, the level of expression of MDR1 gene also will increase. Conclusion: The study results confirm that the the anthracycline drug is one of the MDR1 gene encodes a product specific substrate of p-gp protein. Clinical detection of MDR1 expression levels can be more accurately predict the efficacy of anthracycline and provided the experimental basis for individual choice sensitive drugs. And both anthracyclines (doxorubicin and epirubicin) in vitro induction of MDR1 gene expression was no significant difference in the ability. Meanwhile, the formation of multi-drug resistance in breast cancer is not just based on the selective survival of cancer stem cells may also be partly due to changes in the expression of the adaptive functional genes of breast cancer cells to chemotherapeutic drugs.

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