阿法替尼对卵巢癌阿霉素耐药细胞的增敏作用及其机制

张韫琪; 李国锋; 宋志华

doi:10.3971/j.issn.1000-8578.2018.18.0251

阿法替尼对卵巢癌阿霉素耐药细胞的增敏作用及其机制

Afatinib Sensibilizes Multidrug-resistant Human Ovarian Cancer Cells to Adriamycin and Related Mechanisms

摘要

摘要:
目的探讨阿法替尼增强耐药性卵巢癌细胞对阿霉素化疗敏感度的作用及机制。
方法 MTT法检测联用阿法替尼对阿霉素作用于卵巢癌A2780及A2780T细胞的IC₅₀的影响；不同浓度的阿法替尼干预后，罗丹明123蓄积实验检测ABCB1外排功能；ABCB1-Glo^TM Assay Systems实验检测ABCB1 ATPase活性；Western blot检测A2780T细胞EGFR、p-EGFR、HER-2、p-HER-2及ABCB1的表达；RT-PCR实验检测A2780T细胞MDR1 mRNA的表达。
结果无毒浓度的阿法替尼显著降低了阿霉素对耐药卵巢癌A2780T细胞的IC₅₀（P < 0.05），而对A2780细胞没有影响；阿法替尼浓度依赖性地增加罗丹明123在A2780T细胞内的蓄积量及ABCB1 ATPase活性（P < 0.05）；阿法替尼下调A2780T细胞ABCB1的编码基因MDR1的mRNA水平及ABCB1的蛋白表达水平，抑制了EGFR及HER-2的磷酸化水平。
结论阿法替尼可能通过抑制ABCB1转运体的外排功能，下调ABCB1的表达，进而增敏耐药卵巢癌细胞A2780T对阿霉素的化疗敏感度，是一种有开发前景的卵巢癌化疗增敏剂。

Abstract:
Objective To investigate the effect of afatinib on the chemosensitization of multidrug-resistant A2780T cells to adriamycin and related mechanism.
Methods MTT assay was used to investigate the effect of afatinib on the IC₅₀ values of adriamycin in A2780 and A2780T cells. The efflux function and ATPase activity of ABCB1 were investigated by rhodamine 123 accumulation assay and ABCB1-Glo^TM Assay Systems, respectively. The expression of EGFR, HER-2, p-EGFR, p-HER-2 and ABCB1 were investigated by Western blot. RT-PCR assay was applied to test the mRNA expression of MDR1 in A2780T cells.
Results Afatinib at nontoxic concentrations significantly decreased the IC₅₀ of adriamycin in A2780T cells (P < 0.05), while no effect in A2780 cells. Afatinib could inhibit the efflux function and increase the ATPase activity of ABCB1 in a concentration-dependent manner(P < 0.05). Afatinib inhibited the phosphorylation of EGFR and HER-2, and decreased the expression of ABCB1 protein and MDR1 mRNA in A2780T cells.
Conclusion Afatinib could sensibilize multidrug-resistant A2780T cells to adriamycin by inhibiting the efflux function and downregulating ABCB1 expression.

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