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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A1的K562-pEGFP-N1-ANXA1细胞株。MTT及细胞增殖实验显示,稳定转染膜联蛋白A1的K562-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-N1和K562-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A1与β-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。
Preliminary studies have found that in vitro anti-imatinib-resistant K562 cells, Annexin A1 expression was significantly increased, and increased with the increase in resistance multiple. In order to investigate whether Annexin A1 is involved in the resistance of imatinib, the empty vector control K562-pEGFP-N1 cell line and K562-pEGFP-N1 stably over-expressed Annexin A1 were established by lipofection method. N1-ANXA1 cell line. MTT and cell proliferation experiments showed that K562-pEGFP-N1-ANXA1 stably transfected with annexin A1 increased the sensitivity to imatinib but did not change its proliferation ability. The results of Western blotting showed that K562-pEGFP- There was no significant change in Annexin A1 family proteins, drug-resistance-related proteins and cell proliferation and cell cycle related proteins in N1 and K562-pEGFP-N1-ANXA1 cell lines; co-immunoprecipitation showed that K562-pEGFP-N1-ANXA1 cells The interaction between annexin A1 and β-actin in the strain was significantly enhanced. It is speculated that in vitro established Annexin A1 over-expression of K562 cells, probably due to the high expression of Annexin A1 and its interaction with β-actin promote the absorption of imatinib, thereby increasing K562 cells Sensitivity to imatinib.