非平面多环芳烃[4]helicenium通过促进凋亡选择性杀伤血液恶性肿瘤细胞  

Non-planar PAH [4]helicenium selectively kills hematological malignant tumor cells by promoting apoptosis

作　　者：何小珍 周雁 张元亮[2] 刘培峰[1,3] HE Xiaozhen;ZHOU Yan;ZHANG Yuanliang;LIU Peifeng(Central Laboratory,Ren Ji Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200127,China;State Key Laboratory of Medical Genomics,Shanghai Institute of Hematology,Ruijin Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200025,China;State Key Laboratory of Oncogenes and Related Genes,Shanghai Cancer Institute,Ren Ji Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200032,China)

机构地区：[1]上海交通大学医学院附属仁济医院中心实验室,上海200127 [2]上海交通大学医学院附属瑞金医院,上海血液学研究所,医学基因组学国家重点实验室,上海200025 [3]上海交通大学医学院附属仁济医院,上海市肿瘤研究所,癌基因及相关基因国家重点实验室,上海200032

出　　处：《肿瘤》2020年第9期589-601,共13页Tumor

基　　金：国家自然科学基金资助项目(编号:81771968)。

摘　　要：目的:研究非平面多环芳烃[4]helicenium对血液恶性肿瘤细胞增殖、细胞周期和凋亡的影响,并探讨可能的作用机制。方法:用不同质量浓度的[4]helicenium(0~100μg/mL)分别处理血液恶性肿瘤Kasumi-1、HL-60、K-562、Raji和U-937细胞,采用CCK-8法检测[4]helicenium对5株细胞增殖的影响并计算对各株的半数抑制浓度(half maximal inhibitory concentration,IC50)。以[4]helicenium对Kasumi-1、HL-60、Raji、K-562和U-937细胞各自的IC50为作用浓度处理后,FCM法检测[4]helicenium对细胞周期的影响;实时荧光定量PCR法检测[4]helicenium对5株细胞中细胞周期相关基因细胞周期蛋白A2(cyclin A2)、cyclin B1、cyclin B2,DNA复制相关基因微小染色体维持复合体2(mini-chromosome maintenance complex 2,MCM2)、MCM3、MCM4、MCM6、GINS复合体亚基1(GINS complex subunit 1,GINS1)、GINS4、复制起始复合体亚基5(origin recognition complex subunit 5,ORC5)、核糖核苷酸小亚基M2(ribonucleotide reductase M2,RRM2),DNA修复相关基因范可尼贫血互补群A(Fanconi anemia complementation group A,FANCA)、FANCB、FANCC、FANCD2、FANCE、FANCG、FANCI、FANCL、FA核心复合体相关蛋白100基因(FA core complex associated protein 100,FAAP100)mRNA的表达水平;蛋白质印迹法检测对磷酸化细胞分裂周期2(phospho-cell division cycle 2,p-CDC2)、MCM4、FANCA以及DNA损伤蛋白磷酸化组蛋白H2AX(phospho-histone H2AX,γH2AX)表达水平的影响。FCM法检测[4]helicenium对细胞凋亡的影响。结果:[4]helicenium对Kasumi-1、HL-60、Raji、K-562和U-937细胞的IC50值分别为5.659、5.786、18.700、19.860和26.970μg/mL;[4]helicenium对Kasumi-1和HL-60细胞的抑制作用最明显,其次为Raji和K-562细胞,对U-937细胞的抑制率最低。[4]helicenium可使血液恶性肿瘤细胞周期阻滞在不同阶段,Kasumi-1、HL-60和U-937细胞被阻滞在G2/M期(P值均<0.01),Raji细胞被阻滞在G0/G1期(P<0.05),K-562细胞阻滞在S期(P<0.01)。[4]helicenium可明显下调KasObjective:To investigate the effects of non-planar polycyclic aromatic hydrocarbons[4]helicenium on the proliferation,cell cycle and apoptosis of hematological malignant cells,and to explore the possible mechanism.Methods:Different mass concentrations of[4]helicenium(0-100μg/mL)were used to treat hematological malignant Kasumi-1,HL-60,K-562,Raji and U-937 cells,respectively,The proliferation of Kasumi-1,HL-60,K-562,Raji and U-937 cells was detected by CCK-8 assay and calculated the half maximal inhibitory concentration(IC50).After treatment with IC50 of different cells,FCM assay was used to detect cell cycle.The expression level of cell cycle-related gene cyclin A2,cyclin B1,cyclin B2,DNA replication-related gene mini-chromosome maintenance complex 2(MCM2),MCM3,MCM4,MCM6,GINS complex subunit 1(GINS1),GINS4,origin recognition complex subunit 5(ORC5),ribonucleotide reductase M2(RRM2)and DNA repair-related gene Fanconi anemia complementation group A(FANCA),FANCB,FANCC,FANCD2,FANCE,FANCG,FANCI,FANCL,FA core complex associated protein 100(FAAP100)mRNAs was detected by real-time fluorescent quantitative PCR.The expression level of phospho-cell division cycle 2(p-CDC2),MCM4,FANCA and DNA damage protein phospho-histone H2AX(γH2AX)proteins was detected by Western blotting.Cell apoptosis was detected by FCM assay.Results:The IC50 values of[4]helicenium on Kasumi-1,HL-60,Raji,K-562 and U-937cells were 5.659,5.786,18.700,19.860 and 26.970μg/mL,respectively.[4]Helicenium had the most obvious inhibitory effect on Kasumi-1 and HL-60 cells,Raji and K-562 cells were the second,the inhibitory rate on U-937 cells was the lowest.[4]Helicenium promoted cell cycle arrest in G2/M-phase in Kasumi-1,HL-60 and U-937 cells(all P<0.01),Raji in G0/G1-phase(P<0.05),and K-562 in S-phase(P<0.01).The cell cycle-related gene(cyclin A2,cyclin B1 and cyclin B2),DNA replication-related gene(MCM2,MCM3,MCM4,MCM6,GINS1,GINS4,ORC5,RRM2)and DNA repair-related gene(FANCA,FANCB,FANCC,FANCD2,FANCE,FANCG,FANCI,FANCL,FAAP100)mRNAs as well as p-CDC2,MCM4 an

关 键 词：血液肿瘤 非平面多环芳烃 细胞增殖 细胞凋亡 细胞周期 DNA损伤 

分 类 号：R733[医药卫生—肿瘤]