降低细胞膜角蛋白8和乳腺癌耐药蛋白的表达逆转耐药性  

Reversal multi-drug resistance by reducing the expression of CK8 and BCRP

作　　者：袁翔[1] 程昕[1] 许元富[1] 周圆[1] 邵晓枫[1] 李巍[1] 任思楣[1] 张秀丽[1] 杨铭[1] 

机构地区：[1]中国医学科学院.北京协和医学院血液学研究所,天津300020

出　　处：《中国药理学通报》2009年第11期1425-1429,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30873091);天津市科技计划资助项目(No08ZCGHHZ00900)

摘　　要：目的研究细胞膜角蛋白8(CK8)和乳腺癌耐药蛋白(breast cancer resistant protein,BCRP)作为多靶点治疗逆转耐药的可行性。方法共转染特异的CK8-siRNAs和BCRP-siRNAs至人乳腺癌多药耐药细胞MCF-7/MX,用Western blot方法检测siRNAs对CK8和BCRP蛋白表达的抑制,荧光染色用激光共聚焦显微镜观察细胞膜表面CK8表达量的变化,并用Sulforhodamine B的方法检测转染前后细胞对多种化疗药敏感性的变化。结果MCF-7/MX细胞导入CK8-siRNAs和BCRP-siRNAs后,CK8和BCRP的表达水平均明显降低,且细胞表面CK8染色也明显降低,同时对米托蒽醌等化疗药的敏感性明显提高,耐药表型明显逆转。结论CK8和BCRP在MCF-7/MX耐药细胞中共同高表达且在乳腺癌多药耐药表型的形成中起重要作用,共同抑制CK8和BCRP的表达可以有效逆转乳腺癌的多药耐药,为治疗肿瘤多药耐药提供了一条新的思路。Aim To determine whether membrane cytokeratin 8（CK8 ）and BCRP expression cooperatively contributed to multidrug resistance（MDR）in MCF-7/MX cells.Methods MCF-7/MX cells were transfected with specific anti CK8-siRNAs and anti BCRP-siRNAs via LipofectAMINE2000.The expression of CK8 and BCRP was determined using Western blot,and membrane staining was observed by laser confocal microscopy.Sensitivity to chemical drugs was examined by Sulforhodamine B method.Results The expression levels of cell surface CK8 and BCRP were obviously reduced by siRNAs,and inhibition of CK8 and BCRP expression could effectively restore the sensitivity to drugs and reverse MDR phenotype of MCF-7/MX cells.Conclusions CK8 together with BCRP may play significant roles in conferring the multifactorial MDR phenotype of MCF-7/MX cells,but may act independently via potentially different mechanisms.Combinational approaches that target multiple drug-resistance-related molecules/pathways in cancer cells may represent more efficacious strategies to overcome MDR.

关 键 词：多药耐药 细胞膜角蛋白8 多靶点治疗 乳腺癌耐药蛋白 RNA干扰 米托蒽醌 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R342.2[医药卫生—基础医学]