| 摘要: |
| 目的探讨人乳腺癌MCF-7细胞中第187位苏氨酸（Thr187）磷酸化的细胞周期素依赖性激酶抑制蛋白p27kip1（P-p27Thr187）与化疗药物敏感性的关系。方法人乳腺癌MCF-7细胞常规培养，采用MTT法测定其对5种常用化疗药物[表柔比星（EPI）、多西他赛（DOC）、5-氟尿嘧啶（5-Fu）、环磷酰胺（CTX）、顺铂（CDDP）]的敏感性；采用Westernblot检测经化疗药物作用后P-p27Thr187、p27kip1表达水平；采用免疫共沉淀方法检测P-p27Thr187与S期激酶相关蛋白2（Skp2）相互结合情况。结果5种常用化疗药物对人乳腺癌MCF-7细胞均有抑制作用，5-Fu的抑制率随浓度增加而增高，而其他4种化疗药物的抑制率开始随浓度增加而增高，但达到理论最高药物浓度（PPC）后降低（均P＜0.01）；在PPC下作用48h，抑制率从大到小的化疗药物依次为EPI、DOC、5-Fu、CTX、CDDP。Westernblot结果显示化疗药物作用48h后，P-p27Thr187表达水平明显增加，以EPI、DOC增加最为明显（均P＜0.01）；p27kip1表达水平明显降低（均P＜0.01）。免疫共沉淀结果显示，经EPI0.25滋g/ml、DOC6.80滋g/ml作用48h后，P-p27Thr187与Skp2结合明显增加。结论P-p27Thr187与人乳腺癌MCF-7细胞化疗敏感性有关，可作为预测化疗敏感性及选择药物的一项新指标。 |
| 关键词: 乳腺癌 p27kip1 磷酸化 化疗敏感性 |
| DOI：10.12056/j.issn.1006-2785.2017.39.01.2016-1183 |
| 分类号: |
| 基金项目:浙江省医学重点学科建设计划项目（GJJX-010-001）；嘉兴市科技计划项目（2013AY21042-3）；嘉兴市医学重点支撑学科建设计划项目（04-Z-01）；嘉兴市第一医院“启明星”计划项目（2013QMX003） |
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| Relationship of p27kip1 Thr187 phosphorylation with chemosensitivity of breast cancer MCF-7 cells |
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GUO Zhiqin, LV Qingshan, YUAN Linna, WANG Zhen, LU Ning, KE Ping, WU Wanxin
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Jiaxing First Hospital
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| Abstract: |
| Objective To investigate the relationship of p27kip1 Thr187 phosphorylation(P-p27Thr187) with chemosensitivi- ty of human breast cancer MCF-7 cells. Methods Cultured MCF-7 cells were treated with epirubicin (EPI), docetaxel (DOC), 5-fluorouracil (5-Fu), cyclophosphamide (CTX) and cisplatin (CDDP) , the inhibitory rates were measured by MTT method. The expression of P-p27Thr187 and p27kip1 was detected by Western blot. The conjugation of P-p27Thr187 and S-phase kinase-associated protein 2 (Skp2) was detected by co-immunoprecipitation method. Results The MTT assay showed that inhibitory effect of 5 chemotherapeutical agents on MCF-7 cells in a dose-dependent manner, and with an order of inhibitory effect as: EPI, DOC, 5-Fu, CTX and CDDP. Western blot assay showed that after drugs treatment, the expression of P-p27Thr187 protein was significantly increased, while the expression of p27kip1 was decreased. Co-immunoprecipitation revealed that the conjugation of P-p27Thr187 and Skp2 protein was increased significantly after the chemotherapy drugs treatment. Conclusion P-p27Thr187 protein might be related to the chemosensitivity in human breast cancer MCF-7 cells, and it could be used as a marker for clinical assessment of sensitivity to chemotherapy. |
| Key words: Breast cancer p27kip1 Phosphorylation Chemosensitivity |
