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

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 50 μg/mL与多柔比星联用作用于MCF-7/DOX细胞的IC50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 50 μg/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 50 μg/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。

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

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 CCTα in MCF-7/DOX cells was determined by Western blot. RESULTS: The IC50 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 CCTα was not significantly affected (P>0.05), while the expression of the gene and protein above in OPD 50 μg/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 50 μg/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.