15-PGDH诱导药物逆转人乳腺癌MCF-7/ADR细胞耐药作用及机制

Reversal Effect and Mechanism of 15-PGHD Induction Drugs on the Multidrug Resistance of Human Breast Cancer Cell Line MCF-7/ADR

目的探讨15-羟前列腺素脱氢酶(15-PGDH)在人乳腺癌多药耐药细胞中的表达,并观察15-PGDH诱导药物对MCF-7/ADR细胞耐药性的影响。方法采用逆转录-聚合酶链反应(RT-PCR)、Western blot检测MCF-7和MCF-7/ADR细胞中15-PGDH、环氧合酶-2(COX-2)mRNA和蛋白表达;ELISA检测细胞上清液中PGE_2含量;四甲基偶氮唑蓝(MTT)法测定15-PGDH诱导药物(吲哚美辛、布洛芬、吡格列酮、地塞米松)对乳腺癌细胞的增殖抑制作用及对多柔比星(adriamycin,ADM)敏感性的影响;Hochest 33258检测细胞凋亡。结果与MCF-7相比,MCF-7/ADR细胞中15-PGDH表达水平明显降低,COX-2表达明显增加,细胞上清液中PGE_2水平显著增加;4种15-PGDH诱导药物可增加MCF-7/ADR细胞15-PGDH表达或同时降低COX-2表达,降低细胞上清液中PGE2含量,增强MCF-7/ADR细胞对ADM的敏感性及诱导细胞凋亡的作用,部分逆转耐药。结论15-PGDH在MCF-7/ADR细胞中低表达,其诱导药物可能通过调控15-PGDH和COX-2影响PGE_2表达,增加化疗药物敏感性、促进细胞凋亡、逆转MCF-7/ADR细胞耐药。

OBJECTIVE To examine the expression of 15-hydroxyprostaglandin dehydrogenase（ 15-PGDH） in human multidrugresistant breast cancer line MCF-7/ADR and to explore the reversal effect and mechanism of 15-PGHD induction drugs on MCF-7/ADR cells. METHODS The RT-PCR and Western blot were used to detect 15-PGDH,COX-2 mRNA and protein expression in MCF-7 and MCF-7/ADR cells. PGE_2 levels in supernatant of cells were determined by ELISA assay. Anti-proliferation effect and chemotherapy sensitivity to ADM of 15-PGDH induction drugs（ indomethacin,ibuprofen and pioglitazone,dexamethasone） on breast cancer cells were assayed by MTT method. Cell apoptosis was detected by Hochest 33258 stain assay. RESULTS Compared with MCF-7 cells,the 15-PGDH expression was significantly decreased,COX-2 expression was significantly increased and PGE_2 levels in cell supernatant were increased in MCF-7/ADR cells. 15-PGDH induction drugs（ indomethacin,ibuprofen and pioglitazone,dexamethasone） increased 15-PGDH expression or both reduced COX-2 expression,and finally reduced PGE_2 levels in MCF-7/ADR cells. Effect of chemosensitivity and apoptosis induction of ADM was enhanced and multidrug resistance was partially reversed when co-treated with 15-PGDH induction drugs. CONCLUSION The expression of 15-PGDH is decreased in human multidrug-resistant breast cancer line MCF-7/ADR. 15-PGDH induction drugs could increase chemosensitivity,promote apoptosis and reverse resistance of MCF-7/ADR cell,the mechanism might related to the influence of PGE2 level by regulated the expression of 15-PGDH and COX-2.