摘要
目的探讨抑制C-kit基因突变阳性的t(8;21)急性髓系白血病(acute myeloid leukemia,AML)细胞中缺氧诱导因子1α(hypoxia inducible factor 1α,HIF1α)对C-kit表达及细胞恶性生物学行为的影响。方法采用不同浓度的HIF1α特异性抑制剂棘霉素处理两种携带C-kit基因突变的t(8;21)AML细胞系Kasumi-1和SKNO-124h,以二甲基亚砜处理的细胞作为对照组。采用实时定量RT-PCR检测细胞中HIF1α、C-kit基因mRNA表达水平,Western blot检测蛋白表达水平,AnnexinV-PI双染后流式细胞仪检测细胞凋亡,克隆形成实验检测细胞体外克隆形成能力。结果棘霉素能明显抑制Kasumi-1和SKNO-1细胞中HIF1α基因和C-kit基因mRNA和蛋白的表达水平,且该抑制作用具有浓度依赖性,同时细胞增殖被抑制、细胞凋亡增加。结论棘霉素靶向抑制HIF1α可通过下调C-kit表达抑制携带C-kit基因突变的t(8;21)AML细胞增殖,该作用与棘霉素诱导的细胞凋亡有关。
Objective To investigate the effect of silencing of hypoxia inducible factor 1α(HIF1α)on the expression of C-kit gene and cellular malignant biological behavior in t(8;21)AML carrying C-kit gene mutation.Methods Two t(8;21)AML cell lines carrying C-kit gene mutation,Kasumi-1 and SKNO-1,were respectively treated with different concentration of HIF1αspecific inhibitors echinomycin(1 nmol/L,5 nmol/L,10 nmol/L,20 nmol/L,30 nmol/L,respectively).The cells were harvested at 24 hours after treatment,and those treated with dimethyl sulfoxide were selected as control group.Real-time quantitative RT-PCR was used to detect the HIF1αand C-kit gene mRNA expression levels,Western blot for protein expression levels,AnnexinV-PI double staining followed by flow cytometry for cellular apoptosis,and Colony-forming assay for cellular proliferation in vitro.Results Echinomycinsignificantly inhibited the mRNA and protein expression levels of HIF1αand C-kit genes in Kasumi-1 and SKNO-1 cells with a concentration-dependent manner.The proliferation of both cell lines was inhibited and cellular apoptosis increased.Conclusion Targeted inhibition of HIF1αusing echinomycin can inhibit the proliferation of t(8;21)AML cells carrying C-kit gene mutation by down-regulating the expression of C-kit,which is related to the increased cellular apoptosis caused by echinomycin.
作者
陈泽
张继彬
吕娜
李永辉
周薇
关伟
高晓宁
CHEN Ze;ZHANG Jibin;LYU Na;LI Yonghui;ZHOU Wei;GUAN Wei;GAO Xiaoning(Department of Hematology,Chinese PLA General Hospital,Beijing 100853,China;Department of Cardiology the First Medical Center,Chinese PLA General Hospital,Beijing 100853,China)
出处
《解放军医学院学报》
CAS
2020年第6期608-613,643,共7页
Academic Journal of Chinese PLA Medical School
基金
国家自然科学基金项目(81870109,81670135)
作者简介
陈泽,女,在读硕士。研究方向:白血病的发病机制。Email:cz13021099727@163.com;通信作者:高晓宁,女,副主任医师,副教授,硕士生导师。Email:gaoxn@263.net
