地钱素C对慢性粒细胞白血病K562细胞增殖抑制作用的初步研究  

Preliminary study on the mechanism of proliferation inhibition effect in K562 cells induced by Marchantin C in vitro

作　　者：汪茗[1,2] 席广民 苑辉卿 Wang Ming;Xi Guangmin;Yuan Huiqing(Department of Biochemisty and Molecular Biology,Wannan Medical College,Wuhu Anhui 241002,China;Key Laboratory of Active Macromolecules,Wannan Medical College,Wuhu Anhui 241002,China;Institute of Medical Sciences,the Second Hospital of Shandong University,Jinan Shandong 250014,China)

机构地区：[1]皖南医学院生物化学与分子生物学教研室,安徽芜湖241002 [2]皖南医学院活性大分子省重点实验室,安徽芜湖241002 [3]山东大学第二医院基础医学研究所,山东济南250014

出　　处：《遵义医科大学学报》2022年第6期706-712,共7页Journal of Zunyi Medical University

基　　金：安徽省自然科学基金面上项目(NO:1808085MH272);安徽省高校优秀青年人才支持计划(NO:gxyq2017037)。

摘　　要：目的观察双联苄化合物地钱素C(Marchantin C,MC)对慢性粒细胞白血病K562细胞的增殖抑制作用,初步探讨其抑制K562细胞增殖的作用机制。方法将3、6、12μmol/L的MC作用K562细胞24 h,采用倒置显微镜观察细胞形态学变化,MTT法检测细胞增殖抑制率,DAPI染色观察细胞凋亡,流式细胞术检测细胞凋亡率及周期,Western blotting检测凋亡相关蛋白表达的变化。结果倒置显微镜下MC处理组细胞出现不同程度的分裂受阻、形态不规则,细胞皱缩、核破碎、核溶解等改变。MTT结果显示MC能有效抑制K562细胞增殖。流式细胞术结果显示MC能有效诱导K562细胞凋亡。细胞周期检测结果显示细胞被阻滞在G_(2)/M期。Western blotting结果显示随着MC浓度的增加,剪切形式的Caspase-3表达发生变化,其底物PARP被逐渐剪切,Akt、mTOR、p70S6K的磷酸化形式p-Akt、p-mTOR、p-p70S6K表达有所下调。结论MC能有效抑制K562细胞的增殖,诱导其凋亡,其机制与Akt/mTOR信号通路被抑制有关。Objective To investigate the inhibition effect of Marchantin C(MC) on K562 cells and preliminarily analyze the underlying mechanism.Methods K562 cells were treated with MC(3,6,12 μmol/L for 24 h).Cell morphological changes were oberseved by using inverted microscope.Cell proliferation inhibition rate was measured by MTT assay.Nucleic morphological changes were determined by DAPI staining.The apoptosis rate and cell cycle were detected by flow cytometry(FCM).Western blotting was applied to detect the expression of apoptotic-related proteins in K562 cells.Results Cell division was blocked and cell shrinkage,karyorrhexis could be observed.The proliferation inhibition rate of K562 cells induced by MC was obviously higher than that in control.MC could induce apoptosis of K562 cells.K562 cells were arrested in the G_(2)/M phase.The protein level of cleaved caspase-3 was changed and accompanied with the elevated cleaved PARP.The protein level of phospho-Akt(p-Akt),phospho-mTOR(p-mTOR),phospho-p70 S6 K(p-p70 S6 K) was down-regulated.Conclusion MC can effectively inhibit proliferation and induce apoptosis of K562 cells,which may be mediated via inhibiting the Akt/mTOR/p70 S6 K signaling pathway.

关 键 词：双联苄化合物 白血病 K562细胞 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]