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目的探讨细胞分裂周期蛋白42(Cdc42)在乳腺癌细胞MCF-7阿霉素敏感株和耐药株MCF-7/Adr中表达的变化以及对细胞耐药性的影响。方法将Cdc42 siRNA转染MCF-7/Adr细胞株后,用RT-PCR和Weastern Blot方法检测MCF-7组,MCF-7/Adr组,MCF-7/Adr siRNA干扰组细胞Cdc42的转录以及蛋白的表达水平;采用四甲基偶氮唑蓝(MTT)法测定siRNA处理后阿霉素(ADM)对MCF-7/Adr细胞的杀伤作用;用荧光分光光度计测定细胞内ADM药物浓度。结果 MCF-7/Adr细胞中Cdc42 mRNA及蛋白表达量显著高于MCF-7细胞(P<0.05);siRNA干扰后Cdc42表达受到明显抑制(P<0.05),并显著提高细胞内ADM的浓度(P<0.05)。结论特异性siRNA能明显抑制Cdc42 mR-NA及蛋白表达,并逆转细胞耐药性。
Objective To investigate the expression changes of cell cycle protein 42 (Cdc42) in doxorubicin-resistant MCF-7 and drug-resistant MCF-7 / Adr cells in breast cancer cells and its effect on drug resistance. Methods Cdc42 siRNA was transfected into MCF-7 / Adr cell line, and the transcription of Cdc42 and the protein of MCF-7 / Adr siRNA were detected by RT-PCR and Weastern Blot. The MTT assay was used to determine the cytotoxicity of adriamycin (ADM) on MCF-7 / Adr cells. The intracellular ADM concentration was determined by fluorescence spectrophotometer. Results The expression of Cdc42 mRNA and protein in MCF-7 / Adr cells was significantly higher than that in MCF-7 cells (P <0.05). The expression of Cdc42 was significantly inhibited after siRNA interference (P <0.05) P <0.05). Conclusion Specific siRNA can significantly inhibit the expression of Cdc42 mR-NA and protein, and reverse the cell resistance.