Thr187磷酸化p27^(kip1)与人乳腺癌MCF-7细胞化疗敏感性的关系  

Relationship of p27^(kip1) Thr187 phosphorylation with chemosensitivity of breast cancer MCF-7 cells

作　　者：郭志琴[1] 吕青山 袁琳娜[1] 王振[1] 陆宁[1] 柯萍[1] 邬万新[1] GUO Zhiqin;YUAN Linna;LYU Qingshan

机构地区：[1]嘉兴市第一医院病理科,314000 [2]嘉兴市第一医院检验科,314000

出　　处：《浙江医学》2017年第1期24-27,共4页Zhejiang Medical Journal

基　　金：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)

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p27^(kip1)(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Western blot检测经化疗药物作用后P-p27Thr187、p27^(kip1)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。Western blot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);p27^(kip1)表达水平明显降低(均P<0.01)。免疫共沉淀结果显示,经EPI 0.25μg/ml、DOC 6.80μg/ml作用48h后,P-p27Thr187与Skp2结合明显增加。结论 P-p27 Thr187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。Objective To investigate the relationship of p27kip1 Thr187 phosphorylation(P-p27Thr187) with chemosensitivity 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. Theexpression of P-p27Thr187 and p27kip1 was detected by Western blot. The conjugation of P-p27Thr187 and S-phasekinase-associated protein 2 (Skp2) was detected by co-immunoprecipitation method. Results The MTT assay showed thatinhibitory effect of 5 chemotherapeutical agents on MCF-7 cells in a dose-dependent manner, and with an order of inhibitoryeffect as: EPI, DOC, 5-Fu, CTX and CDDP. Western blot assay showed that after drugs treatment, the expression of P-p27Thr187protein was significantly increased, while the expression of p27kip1 was decreased. Co-immunoprecipitation revealed that theconjugation 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.

关 键 词：乳腺癌 P27KIP1 磷酸化 化疗敏感性 

分 类 号：R737.9[医药卫生—肿瘤]