氧化前胡素对乳腺癌MCF-7/DOX细胞多柔比星耐药的抑制作用

Effects of oxypeucedanin on the resistance of breast cancer MCF-7/DOX cells to doxorubicin

目的:探讨氧化前胡素(oxypeucedanin,OPD)对人乳腺癌MCF-7/DOX细胞多柔比星耐药的影响并探究其可能机制。方法:体外培养MCF-7/DOX细胞,MTT法检测OPD对MCF-7/DOX细胞增殖的影响,并考察OPD最大无毒浓度与多柔比星不同浓度联用对MCF-7/DOX细胞增殖的影响。qRT-PCR检测OPD与多柔比星联用对MCF-7/DOX细胞MDR1、MRP1及甘油磷脂代谢酶AGPAT2、CHKA、CEPT1、DGKA、PCYT1A、PLA2G15 mRNA水平的影响。Western blot检测OPD与多柔比星联用对MCF-7/DOX细胞MDR1、MRP1、CHKA及CCTα蛋白表达的影响。结果:OPD 50μg/mL与多柔比星联用作用于MCF-7/DOX细胞的IC_(50)值低于多柔比星单独,其逆转倍数为1.56倍。与对照组比较,多柔比星组MCF-7/DOX细胞MDR1、MRP1和6种甘油磷脂代谢酶mRNA均下调(P<0.05或P<0.01),MDR1、MRP1、CHKA和CCTα蛋白表达无显著变化(P>0.05),OPD 50μg/mL与多柔比星联用组上述基因和蛋白表达均显著下调(P<0.05)。与多柔比星组比较,OPD 50μg/mL与多柔比星联用组MCF-7/DOX细胞上述基因表达进一步下调(P<0.05或P<0.01),上述4种蛋白表达无显著变化(P>0.05)。结论:OPD可增强MCF-7/DOX细胞对多柔比星的化疗敏感性,逆转耐药,可能与其抑制甘油磷脂代谢途径有关。

AIM:To investigate the effect of oxypeucedanin(OPD)on doxorubicin resistance in human breast cancer MCF-7/DOX cells and its possible mechanism.METHODS:MCF-7/DOX cells were cultured in vitro,MTT assay was used to detect the effect of OPD on the survival of MCF-7/DOX cells,and the effect of OPD combined with different concentrations of doxorubicin on the proliferation of MCF-7/DOX cells were investigated.The effect of OPD combined with doxorubicin on the expression of genes including MDR1,MRP1,AGPAT2,CHKA,CEPT1,DGKA,PCYT1A,PLA2G15 in MCF-7/DOX cells was measured by qRT-PCR.The effect of OPD combined with doxorubicin on the protein expression of MDR1,MRP1,CHKA and CCTαin MCF-7/DOX cells was determined by Western blot.RESULTS:The IC_(50) value of doxorubicin in combination with OPD in MCF-7/DOX cells was lower than that of doxorubicin alone.The reversal fold is 1.56.Compared with control group,the mRNA level of MDR1,MRP1,AGPAT2,CHKA,CEPT1,DGKA,PCYT1A and PLA2G15 in the doxorubicin group was significantly downregulated(P<0.05 or P<0.01),the protein expression of MDR1,MRP1,CHKA and CCTαwas not significantly affected(P>0.05),while the expression of the gene and protein above in OPD 50μg/mL combined with doxorubicin group was significantly downregulated(P<0.05 or P<0.01).Compared with doxorubicin group,the expression of the gene above in OPD 50μg/mL combined with doxorubicin group was further downregulated(P<0.05 or P<0.01),but the expression of the protein above was not significantly affected(P>0.05).CONCLUSION:OPD can enhance the sensitivity of MCF-7/DOX cells to doxorubicin chemotherapy,reverse drug resistance,which may be related to inhibition of glycerophospholipid metabolism.