耐药逆转剂对人乳腺癌细胞P-糖蛋白、EMMPRIN和MMP表达的影响

Effects of multidrug resistance reversing agent on P-glycoprotein,EMMPRIN and MMP expression in human breast cancer cells

目的观察耐药逆转剂川芎嗪(tetramethylpyrazine,TMP)对人乳腺癌MCF7/Adr细胞P-糖蛋白(P-glycoprotein,P-gp)、基质金属蛋白酶诱导物(extracellular matrix metalloproteinase inducer,EMMPRIN)和基质金属蛋白酶2(matrix metalloproteinases 2,MMP2)、MMP9表达的影响。方法采用四甲基偶氮唑盐(MTT)比色法检测药物对MCF7/Adr细胞的毒性作用;荧光分光光度法测定细胞内阿霉素(adriamycin,ADM)的荧光强度;Real time RT-PCR和Western blot检测细胞P-gp、EMMPRIN、MMP2和MMP9 mRNA和蛋白水平的变化。结果TMP与ADM联合作用于细胞,与单用ADM相比,ADM对细胞的杀伤效应及细胞内ADM的荧光强度明显升高(P<0.05),且有浓度依赖性;与对照组相比,ADM能上调细胞P-gp、EMMPRIN和MMP2、MMP9蛋白表达;TMP能抑制ADM对细胞P-gp、EMMPRIN、MMP2和MMP9蛋白和mRNA水平的上调作用。结论TMP在有效逆转肿瘤多药耐药的同时,还能抑制ADM对P-gp、EMMPRIN、MMP2和MMP9的上调。

Objective To investigate the effects of multidrug resistance （MDR） reversing agent tetramethylpyrazine （TMP） on P-glycoprotein（P-gp）, EMMPRIN, MMP2 and MMP9 expression in human breast cancer cells. Methods The toxicity of the drugs to MCF7/Adr cells was assessed by MTT colorimetric assay; The fluorescence intensity of adriamycin （ADM） in MCF7/Adr cells was detected with fluorescence spectrophotometer; Expression of P-gp, EMMPRIN, MMP2 and MMP9 was detected by Western blot analysis and reverse transcription and quantitative real-time polymerase chain reaction （Real time RT-PCR）. Results The combination of TMP and ADM enhanced the cytotoxicity of ADM and elevated the fluorescence intensity of ADM in MCF7/Adr cells as compared with the ADM group （P〈0.05）, and these effects were dose-dependent. Compared with the control group, P-gp, EMMPRIN, MMP2 and MMP9 were up-regulated by ADM at expression level and the up-regulation was inhibited by TMP at both mRNA and protein levels. Conclusions TMP not only reversed MDR effectively, but also inhibited the up-regulation of P-gp, EMMPRIN, MMP2 and MMP9 by ADM.