阳离子脂质体-siRNA复合物对K562细胞凋亡诱导的作用机制  

Mechanism of cationic liposome-siRNA complex on apoptosis induction of K562 cells

作　　者：王保全 陈江涛 申智慧[3] 周黎明[4] 李文倩 史朝辉 翟巧玲 张永州 WANG Baoquan;CHEN Jiangtao;SHEN Zhihui;ZHOU Liming;LI Wenqian;SHI Zhaohui;ZHAI Qiaoling;ZHANG Yongzhou(Department of Pharmacy,Huaihe Hospital of Henan University,Henan,Kaifeng 475004,China;Department of General Surgery,Huaihe Hospital of Henan University,Henan,Kaifeng 475004,China;Department of Hematology,Henan Cancer Hospital,Zhengzhou 450009,China;Department of Pharmacy,Zhengzhou Yihe Hospital,Henan University,Zhengzhou 450047,China)

机构地区：[1]河南大学淮河医院药学部,河南开封475004 [2]河南大学淮河医院普外科,河南开封475004 [3]河南省肿瘤医院血液科,河南郑州450009 [4]河南大学郑州颐和医院药学部,河南郑州450047

出　　处：《中国现代医生》2022年第25期31-35,共5页China Modern Doctor

基　　金：河南省科技厅项目(182102311244)。

摘　　要：目的制备新型阳离子脂质体-siRNA复合物,并探讨其对K562细胞凋亡的诱导作用及作用机制。方法通过逆向蒸发技术制备阳离子脂质体复合物,以K562细胞为模型,使用不同浓度的脂质体-siRNA复合物转染K562细胞,MTT法测定脂质体-siRNA复合物对K562细胞生长的影响,流式细胞术检测脂质体-siRNA复合物对K562细胞周期的影响,反转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)和蛋白质印迹法检测脂质体-siRNA复合物对K562细胞相关mRNA及蛋白表达的影响。结果与对照组相比,脂质体-siRNA复合物转染K562细胞增殖受到明显抑制,流式细胞术检测结果显示,阻滞于G_(1)期细胞明显增多(P<0.05),而S期和G_(2)/M期细胞显著减少(P<0.05),RT-PCR和蛋白质印迹法结果显示,随着脂质体-siRNA复合物浓度的增加,K562细胞中bcl-2蛋白及mRNA表达水平均显著降低(P<0.05),bax、caspase-3蛋白及mRNA表达水平均显著升高(P<0.05)。结论新型脂质体-siRNA复合物转染率高,且沉默效果显著,可下调bcl-2基因,上调bax、caspase-3基因,进而促进K562细胞凋亡,有望成为慢性粒细胞白血病基因治疗的高效药物传递系统。Objective To prepare a novel cationic liposome-siRNA complex,and to explore the apoptotic induction of K562 cells induced by liposome-siRNA complex and its mechanism.Methods Cationic liposome complex was prepared by reverse evaporation.K562 cells were used as a model and transfected with different concentrations of liposome-siRNA complex,MTT assay was used to detect the effect of liposome-siRNA complex on K562 cell proliferation.Flow cytometry was used to detect the effect of liposome-siRNA complex on K562 cell cycle.The mRNA and protein expression of bcl-2,bax and caspase-3 in K562 cells were detected by reverse transcription polymerase chain reaction(RT-PCR)and Western blotting.Results Compared with control group,the proliferation of K562 cells transfected with liposome-siRNA complex was significantly inhibited,and the flow cytometry results showed that K562 cells in G_(1)phase were significantly inceased(P<0.05),while cells in S phase and G_(2)/M phase were significantly decreased(P<0.05).RT-PCR and Western blotting results showed that the expression levels of bcl-2 protein and mRNA were significantly decreased(P<0.05),the expression levels of bax、caspase-3 protein and mRNA were significantly increased(P<0.05),with the increase of the concentration of liposome-siRNA complex.Conclusion The novel liposome-siRNA complex has high transfection rate and significant silencing effect,and can down-regulate bcl-2 gene,up-regulate bax and caspase-3 genes,and promote K562 cell apoptosis,which is expected to become an efficient drug delivery system for gene therapy of chronic melogenous leukemia.

关 键 词：脂质体-siRNA复合物 K562细胞 慢性粒细胞白血病 凋亡诱导 

分 类 号：R91[医药卫生—药学]