不同性激素对P-糖蛋白表达水平及甲磺酸伊马替尼转运的影响

Effects of different sex hormones on P-gp expression and imatinib mesylate transport

目的:探讨在ABCB1(1199G/A)重组细胞模型中不同性激素对P-糖蛋白(P-gp)表达水平及甲磺酸伊马替尼转运的影响。方法:将构建的ABCB1_(1199G/wt)和ABCB1_(1199A/mut)重组质粒稳定转染入HEK293细胞中,采用RT-PCR和Western Blot分别检测不同性激素对P-gp mRNA和蛋白的表达水平;采用HPLC检测性激素对甲磺酸伊马替尼累积量的影响。结果:浓度梯度的性激素(雌酮、雌三醇雌二酮、黄体酮和睾酮)刺激ABCB1_(1199G/wt)和ABCB1_(1199A/mut)重组细胞后,雌酮和雌三醇能刺激P-gp mRNA和蛋白表达水平,且雌三醇上调ABCB1_(1199A/mut)mRNA的表达水平显著高于ABCB1_(1199G/wt);雌酮和雌三醇可降低甲磺酸伊马替尼在细胞内的累积量,产生对伊马替尼的耐药性。结论:雌酮和雌三醇增加了细胞内P-gp的表达,并促进P-gp介导甲磺酸伊马替尼的转运能力。

OBJECTIVE To investigate the effect of different sex hormones on the expression of P-gp and the transport of imatinib mesylate in ABCB1（1199 G/A）recombinant cell model.METHODS The recombinant plasmids of ABCB1（1199 G/A）were stably transfected into HEK293 cells,and the expression of P-gp mRNA and protein was detected by RT-PCR and Western blot,respectively.The cumulative amount of imatinib was detected by HPLC.RESULTS The recombinant ABCB1 1199 G/wt and ABCB1 1199A/mut cells were incubated with various concentrations of hormones（estrone,estradiol,estriol,progesterone and testosterone）,the results of Western blot and RT-PCR showed that the expressions of P-gp protein and mRNA were stimulated by estrone or estriol.Moreover,the up-regulation of mRNA expression in ABCB1 1199 A/mut cells by estriol was significantly higher than ABCB1 1199 G/wt cells.Estrone and estriol could reduce the intracellular accumulation of imatinib mesylate.CONCLUSION Estrone or estriol of certain concentrations can up-regulate the expression levels of P-gp and mRNA.Estrone and estriol can reduce the intracellular accumulation of imatinib mesylate,and promote the transport ability of P-gp-mediated imatinib mesylate.