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目的采用比较蛋白质组学的方法,检测多柔比星对乳腺癌细胞核内丝切蛋白(cofilin)相互作用蛋白的影响。方法采用Western blot检测多柔比星对细胞核内cofilin表达的影响;免疫共沉淀后,采用比较蛋白质组学方法,检测对照组与多柔比星处理组细胞核内cofilin相互作用蛋白的差异;采用流式细胞仪检测多柔比星对细胞周期的影响。结果多柔比星处理MDA-MB-231细胞后,细胞核内cofilin表达增加,呈明显量效和时效关系;与对照组比较,多柔比星处理组核内与cofilin相互作用的蛋白结合蛋白、结构分子激活蛋白、细胞骨架蛋白、核酸结合蛋白、核糖体蛋白的种类明显减少(109、60、45、76、34 vs 79、32、20、56、16);多柔比星处理细胞后,可引起细胞周期阻滞在S/G2期,并呈明显量效关系,G1期细胞由(45.92±4.57)%降至(10.50±0.83)%(P<0.01)。结论多柔比星处理细胞后可增加cofilin核聚集,减少细胞核内与cofilin相互作用蛋白的种类,最终使细胞周期阻滞在S/G2期。
Objective To compare the effect of doxorubicin on cofilin-interacting protein in breast cancer cells by comparing proteomics methods. Methods Western blotting was used to detect the effect of doxorubicin on the expression of cofilin in the nucleus. After co-immunoprecipitation, the differences of cofilin-interacting protein in control group and doxorubicin-treated group were detected by comparative proteomics. Cytometry detects the effect of doxorubicin on cell cycle. Results After treated with doxorubicin, the expression of cofilin in nucleus increased after MDA-MB-231 cells were treated with doxorubicin. Compared with the control group, the protein-binding protein interacting with cofilin in doxorubicin- The structure of molecular activator protein, cytoskeleton protein, nucleic acid binding protein, ribosomal protein was significantly reduced (109,60,45,76,34 vs 79,32,20,56,16); doxorubicin treatment of cells, The cell cycle arrest in S / G2 phase was significantly dose-effect relationship. The percentage of cells in G1 phase decreased from (45.92 ± 4.57)% to (10.50 ± 0.83)% (P <0.01). Conclusion Doxorubicin can increase cofilin nuclear aggregation and decrease the types of proteins interacting with cofilin in the nucleus, eventually leading to cell cycle arrest in S / G2 phase.