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400038 重庆,第三军医大学药学系生药学与天然药物化学教研室
[高宁,E-mail: gaoning59@163.com Gao Ning, E-mail: gaoning59@163.com
收稿:2016-11-03,
修回:2016-12-18,
纸质出版:2017-06
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刘艳霞, 李雨浓, 傅若秋, 等. 多柔比星对乳腺癌细胞核内丝切蛋白相互作用蛋白影响的比较蛋白质组学研究[J]. 第三军医大学学报, 2017,39(6):522-528.
Yanxia Liu, Yunong Li, Ruoqiu Fu, et al. Effects of doxorubicin on cofilin interacting proteins in nucleus of breast cancer cells through comparative proteomics[J]. Journal of Third Military Medical University, 2017, 39(6): 522-528.
刘艳霞, 李雨浓, 傅若秋, 等. 多柔比星对乳腺癌细胞核内丝切蛋白相互作用蛋白影响的比较蛋白质组学研究[J]. 第三军医大学学报, 2017,39(6):522-528. DOI: 10.16016/j.1000-5404.201611024.
Yanxia Liu, Yunong Li, Ruoqiu Fu, et al. Effects of doxorubicin on cofilin interacting proteins in nucleus of breast cancer cells through comparative proteomics[J]. Journal of Third Military Medical University, 2017, 39(6): 522-528. DOI: 10.16016/j.1000-5404.201611024.
目的
2
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
2
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
2
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/G
2
期,并呈明显量效关系,G
1
期细胞由 (45.92±4.57)%降至 (10.50±0.83)% (
P
<
0.01)。
结论
2
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/G
2
期。
Objective
2
To determine the effects of doxorubicin on cofilin interacting proteins in the nucleus of breast cancer cells.
Methods
2
The expression of cofilin in the nucleus was evaluated by Western blotting. After immunoprecipitation
the differences of cofilin interacting proteins in the nucleus between the controls and doxorubicin-treated cells were determined by comparative proteomics. The effects of doxorubicin on cell cycle were determined by flow cytometry.
Results
2
Doxorubicin treatment resulted in an increase of cofilin level in the nucleus of MDA-MB-231 cells in dose-and time-dependent manner. Compared with the controls
doxorubicin treatment also resulted in a decreases of the types of cofilin interacting proteins including protein binding proteins
structural molecule activity proteins
cytos
keleton proteins
nucleic acid binding proteins
and ribosome proteins (109
60
45
76
and 34
vs
79
32
20
56
and 16
respectively). Furthermore
doxorubicin treatment led to cell cycle arrest at S/G
2
phase in a dose-dependent manner and the percentage of cells at G
1
phase was declined from (45.92±4.57)% to (10.50±0.83)% (
P
<
0.01).
Conclusion
2
Doxorubicin treatment increases the accumulation of cofilin in the nuclear
decreases the types of cofilin interacting proteins
and finally leads to cell cycle arrest at S/G2 phase.
李朵璐, 周玉冰, 韩超, 转录信号转导子与激活子3小分子干扰RNA增强乳腺癌对多柔比星的药物敏感性[J].中国药学杂志, 2016, 51(11): 896-903. DOI: 10. 11669 /cpj. 2016. 11. 007
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章常华, 涂秀英, 熊淑平, 等.基于蛋白质组学技术筛选葛根芩连汤防治胰岛素抵抗作用的血清差异表达蛋白[J].中药新药与临床药理, 2016, 27(1): 81-85. DOI: 10.3969/j.issn.1003-9783.2016.01.018
Zhang C H, Tu X Y, Xiong S P, et al. Screening for serum differentially-expressed proteins Gegen Qin Lian decoctionin treating insulin resistance with proteomics technology[J ] . Traditional Chinese Drug Research & Clinical Pharmacology, 2016, 27(1): 81-85. DOI: 10.3969/j.issn.1003-9783.2016.01.018
Grzanka D, Marszalek A, Izdebska M, et al. Actin cytoskeleton reorganization correlates with cofilin nuclear expression and ultrastructural changes in cho aa8 cell line after apoptosis and mitotic catastropheinduction by doxorubicin[J]. Ultrastruct Pathol, 2011, 35(3): 130-138. DOI: 10.3109/01913123.2010.548113
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