药学学报, 2013, 48(6): 866-873
引用本文:
李康宁, 金 晶, 陈晓光. 膜联蛋白A1增加人慢性髓系白血病细胞K562对伊马替尼的敏感性[J]. 药学学报, 2013, 48(6): 866-873.
LI Kang-ning, JIN Jing, CHEN Xiao-guang. Annexin A1 increases the sensitivity of K562 cell to imatinib[J]. Acta Pharmaceutica Sinica, 2013, 48(6): 866-873.

膜联蛋白A1增加人慢性髓系白血病细胞K562对伊马替尼的敏感性
李康宁, 金 晶, 陈晓光*
(中国医学科学院、北京协和医学院药物研究所, 天然药物活性物质与功能国家重点实验室, 北京 100050)
摘要:

前期研究发现, 体外建立的耐伊马替尼的K562细胞中, 膜联蛋白A1的表达明显上调, 且随耐药倍 数的增加而增加。为了研究膜联蛋白A1是否参与了伊马替尼的耐药, 采用脂质体转染的方法建立了空载体对照K562-pEGFP-N1细胞株和稳定过表达膜联蛋白A1K562-pEGFP-N1-ANXA1细胞株。MTT及细胞增殖实验显, 稳定转染膜联蛋白A1K562-pEGFP-N1-ANXA1细胞株对伊马替尼的敏感性增加, 但增殖能力不变; Western blotting检测结果显示, K562-pEGFP-N1K562-pEGFP-N1-ANXA1细胞株中的膜联蛋白A1家族蛋白、耐药相关蛋白以及细胞增殖和周期相关蛋白均无明显改变; 免疫共沉淀结果显示, K562-pEGFP-N1-ANXA1细胞株中膜联蛋白A1β-actin的相互作用明显增强。以此推测, 在体外建立的膜联蛋白A1过表达的K562细胞中, 可能由于膜联蛋白A1的高表达及其与β-actin的相互作用促进了伊马替尼的吸收, 从而增加K562细胞对伊马替尼的敏感性。

关键词:   
Annexin A1 increases the sensitivity of K562 cell to imatinib
Abstract:

Annexin A1 (ANXA1) is a kind of endogenous scaffold protein.  Previous research showed that ANXA1 could increase markedly with multiple increase of drug resistance in K562/imatinib cell lines in vitro.  Here the stable transfection cell strains K562-pEGFP-N1 which was the native control and K562-pEGFP-N1- ANXA1 which can stably express ANXA1 were established using the Lipofectamine 2000 in order to find whether ANXA1 involved in the drug resistance.  Cell growth inhibition experiment via MTT and cell proliferation experiment via MTS showed that K562-pEGFP-N1-ANXA1 cell strain was more sensitive to imatinib than the K562-pEGFP-N1 cell strain, and however the ability of proliferation of K562-pEGFP-N1-ANXA1 cell strain did not change compared with the negative control.  Western blotting results showed that the expression of proteins in Annexin family did not change; drug resistance proteins, Bcr-Abl/p-Bcr-Abl (Tyr245), Src family kinase for example, did not change; proteins related with cell proliferation and cell cycle, such as ERK1/2MAPK, p-38MAPK, CDK1 and Wee 1, did not change either in the K562-pEGFP-N1-ANXA1 cell strain compared with the negative control.  The co-immunoprecipitation result showed that the interaction between ANXA1 and β-actin in the K562-pEGFP-N1-ANXA1 cell strain increased markedly.  The deduction was that ANXA1 may make the K562-pEGFP-N1-ANXA1 cell strain more sensitive to imatinib due to the increased uptake of imatinib via the increase of ANXA1 and the interaction between ANXA1 and β-actin in the K562-pEGFP-N1-ANXA1 cell strain in vitro.

Key words:   
收稿日期:
相关功能
PDF(4816KB) Free
打印本文
0
作者相关文章