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Cytoplasmic Macrophage Colony-stimulating Factor Induces Adriamycin-resistance by Up-regulating Mdr-1 Expression and Pumping Out of the Drug Mediated Via PI3K Signaling in Human Breast Cancer MCF-7 Cells.

Author: LiuShaoZhi
Tutor: TangShengSong;WangZuoHua
School: Nanhua University
Course: Pharmacology
Keywords: Maerophage colony-stimulatingacftor (M-CSF) human breast cancer MCF-7 cell line Multidrugresistance P-gp
CLC: R737.9
Type: Master's thesis
Year: 2011
Downloads: 36
Quote: 0
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Abstract


OBJECTIVES:To explore the mechanism and effects of cytoplasmic M-CSF on the sensitivity of adriamycin in human breast cancer MCF-7 cells.METHODS: MCF-7 cell, MCF-7-C cell and MCF-7-M cell were treated with adriamycin at a concentration of 0.05、0.5 and 5.0μg/mL following with or without pretreatment of a specific inhibitor of PI3K LY294002. The effect of cytoplasmic M-CSF on the sensitivity of MCF-7 cells were analyzed by MTT. Protein expression level of p-gp and PKCαwas assessed by Western blotting. Mdr-1 expression was detected by RT-PCR.High efficiency fluid chromatography(HPLC) was used to measure the drug concentration within the cells.RESULTS:The results from Western blotting indicate that expression of P-gp protein is significantly higher in MCF-7-M cells than that in either MCF-7-C cells or MCF-7 cells(p<0.05).LY294002,a specific inhibitor of PI3K could remarkably decreased the expression of P-gp in MCF-7-M cells, compared with either MCF-7-C cells or MCF-7 cells (p<0.05). There is little difference for expression of PKCαbetween MCF-7-M cells and MCF-7 cell or MCF-7-C(p>0.05). The results from RT-PCR show that expression of Mdr-1 is markedly higher in MCF-7-M cells than that in either MCF-7-C cells or MCF-7 cells(p<0.05). and PI3K inhibitor LY294002 remarkably decreases the expression of Mdr-1 in MCF-7-M cells (p<0.05).The results from HPLC show that the concentration of adriamycin within MCF-7-M cells is significantly than that within either MCF-7-C or MCF-7(p<0.05). PI3K inhibitor LY294002 remarkably increases the concentration of adriamycin within MCF-7-M cells (p<0.05). The results from MTT show the sensitivity of adriamycin on MCF-7-M cells is markedly lower than that of either MCF-7-C cells or MCF-7 cells(p<0.05),The IC50 of MCF-7-M cell, MCF-7-C cell and MCF-7 cell is 237.04, 19.26,21.70,repectively.LY294002 could increases c the sensitivity of adriamycin on MCF-7-M cells (p<0.05), the IC50 of MCF-7-M cell, MCF-7-C cell and MCF-7 cell is 83.41,30.16 ,28.33, repectively.CONCLUSION : Cytoplasmic M-CSF induces the adriamycin-resistance and expression of both P-gp and mdr-1. Cytoplasmic M-CSF induces adriamycin-resistance byupregulating the expression of mdr-1 and pumping out of the drug mediated via PI3K signaling in human breast cancer MCF-7 cells.

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