CMPK1蛋白与阿霉素引起的多药耐药的相关性研究

Relationship between CMPK1 protein and ADM caused multidrug resistance

目的对阿霉素可能的作用靶点进行分析,筛选阿霉素耐药相关蛋白。方法质粒转染HEK239细胞以构建蛋白高表达模型;IC_(50)检测细胞的药敏性;RT-PCR检测细胞内基因的表达;Western blot检测CMPK1蛋白的表达;CMPK1siRNA构建CMPK1蛋白低表达模型。结果 IC_(50)检测结果表明高表达CMPK1蛋白的细胞对阿霉素的敏感性增加程度最大(IC_(50)^(HEK293-CMPK1)/IC_(50)^(HEK293-Control)=0.15,P<0.01),且阿霉素耐药细胞(MCF7/ADM)中CMPK1蛋白的表达低于乳腺癌亲本细胞MCF7(P<0.05)。MCF7细胞中,CMPK1蛋白表达水平经CMPK1 siRNA下调之后,对阿霉素的药敏性随之降低(IC_(50)^(MCF7-siCMPK1)/IC_(50)^(MCF7-Control)=3.6,P<0.01),且对紫杉醇、吉西他滨的药敏性也随之降低。结论 CMPK1与细胞的多药耐药相关,且CMPK1蛋白的表达与药敏性呈正相关,提示了CMPK1作为多药耐药治疗靶点的可能性。

Aim To assay the possible targets of adria- mycin （ ADM） , screening ADM resistance related pro-teins. Methods The drug sensitivity of the cells was analyzed by IC50 assay; RT-PCR assay was used to de-tect the expression of genes in the cells; CMPK1 pro-tein expression was tested by Western blot assay; the expression of CMPK1 in the cells was decreased by siRNA of CMPK1. Results Data from IC50 assay showed the sensitivity of cells transfected with CMPK1 was increased most （ IC50 HEK293-CMPK /IC50 HEK293-Control = 0. 15 , P 〈 0. 01 ） , and the expres-sion of CMPK1 protein in ADM resistant breast cells （MCF7/ADM） was lower than that in parent MCF7cells （ P 〈 0.05 ） . When the expression level of CMPK1 was decreased by CMPK1 siRNA, the sensitiv-ity of MCF7 cells to ADM decreased （ IC 50 MCF7-siC- MPK1/IC50MCF7-Control = 3. 6 , P〈 0 .0 1 ） , and the sensitivity of MCF7 cells to paclitaxel and gemcitabine also decreased. Conclusions CMPK1 was related to the multidrug resistance of cells, and the expression of CMPK1 was positively related to the sensitivity to drugs, which provides the possibility of CMPK1 as a target in the treatment of multidrug resistance.