芪贞归脾汤对淋巴瘤细胞株生物学行为的影响及其机制探讨  

Effect of Qi-Zhen-Gui-Pi decoction on biological behavior of lymphoma cells and its mechanism

作　　者：张影 路莎莎[1] 蔡佳翌[1] 李鹤[2] 黄洪晖[1] ZHANG Ying;LU Shasha;CAI Jiayi;LI He;HUANG Honghui(Department of Hematology,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China;Department of Traditional Chinese Medicine,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China)

机构地区：[1]上海交通大学医学院附属仁济医院血液科,上海200127 [2]上海交通大学医学院附属仁济医院中医科,上海200127

出　　处：《肿瘤》2019年第5期346-358,共13页Tumor

基　　金：上海市进一步加快中医药事业发展三年行动计划(2014年-2016年)(编号:ZY3-CCCX-3-3037);上海市浦东新区卫生和计划生育委员会科技发展专项(编号:PW2015E-1)~~

摘　　要：目的 :探讨芪贞归脾汤对淋巴瘤细胞增殖、凋亡、细胞周期、迁移和侵袭的影响,并通过高通量组学技术探索其可能的分子作用机制。方法:采用不同浓度的芪贞归脾汤作用于淋巴瘤Jurkat、SU-DHL-4和Raji细胞株;CCK-8法检测细胞的增殖抑制率;FCM法检测细胞凋亡率及细胞周期;Transwell小室法检测细胞的迁移和侵袭能力;采用PrimeView人类基因表达谱芯片(PrimeView Human Gene Expression Array)检测基因表达谱变化;应用IngenuityPathwayAnalysis(IPA)进行经典通路分析;选择有差异表达的6个基因[磷脂酰肌醇3-激酶催化亚单位α(phosphatidylinositol3-kinasecatalyticsubunitalpha,PIK 3CA)、B细胞κ轻肽基因增强子核因子1(nuclearfactorofkappalightpolypeptide geneenhancerinB-cells1,NFKB 1)、B细胞κ轻肽基因增强子抑制因子,激酶β(inhibitor of kappa light polypeptide gene enhancer in B-cells,kinase beta,IKBKB)、B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、Janus激酶2(Januskinase2,JAK 2)和信号转导及转录激活因子3(signaltransducer and activator of transcription 3,STAT 3)]进行实时荧光定量PCR法验证。结果 :质量浓度为5～25 mg/mL的芪贞归脾汤对Jurkat、SU-DHL-4和Raji细胞株均有明显的增殖抑制作用(P值均<0.01)。芪贞归脾汤质量浓度为20和25 mg/mL组Jurkat及Raji细胞的凋亡率显著升高(P值均<0.01)。10、15和20mg/mL芪贞归脾汤作用48h后,3株淋巴瘤细胞的细胞周期均被阻滞在G2/M期(P值均<0.01)。15mg/mL芪贞归脾汤可显著抑制Jurkat及Raji细胞的迁移和侵袭能力(P值均<0.05)。Jurkat细胞在15mg/mL芪贞归脾汤作用48h后,共有648个基因表达上调和1022个基因表达下调。基于IPA的经典通路分析结果显示,差异基因显著富集的信号通路中磷脂酰肌醇3-激酶(phosphoinositide3-kinase,PI3K)-蛋白激酶B(protein kinaseB,PKB,又称Akt)信号通路(P <0.001)及JAK/STAT信号通路(P <0.05)明显受到抑制。PIK3CA、NFKB1、IKBKB、Bcl-2、JAK2和STAT3 mObjective: To investigate the effects of Qi-Zhen-Gui-Pi decoction on the proliferation, apoptosis, cell cycle, migration and invasion of lymphoma cells, and to explore the possible molecular mechanism using high throughput omics technology. Methods: The human lymphoma cell lines(Jurkat, SU-DHL-4, and Raji) were treated with different concentrations of Qi-Zhen-Gui-Pi decoction. The proliferation inhibitory rate of lymphoma cells was detected by CCK-8 method. The changes of apoptosis and cell cycle distribution were detected by FCM assay. The migration and invasion abilities of lymphoma cells were detected by Transwell chamber assay. The gene expression profiles were detected by PrimeView Human Gene Expression Array. Ingenuity Pathway Analysis(IPA) was used to analyze the classical signaling pathway. The expression levels of differentially expressed genes including phosphatidylinositol 3-kinase catalytic subunit alpha(PIK 3 CA), nuclear factor of kappa light polypeptide gene enhancer in B-cells 1(NFKB 1), inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta(IKBKB), B-cell lymphoma-2(Bcl-2), Janus kinase 2(JAK2) and signal transducer and activator of transcription 3(STAT3) were detected by real-time fluorescent quantitative PCR.Results: After the treatment with different concentrations of Qi-Zhen-Gui-Pi decoction(5-25 mg/mL), the proliferation of Jurkat, SU-DHL-4 and Raji cells was significantly inhibited(all P<0.01). After the treatment with 20 mg/mL and 25 mg/mL Qi-Zhen-Gui-Pi decoction, the apoptosis rates of Jurkat and Raji cells were significantly increased(all P<0.01). After the treatment with different concentrations of Qi-Zhen-Gui-Pi decoction(10, 15 and 20 mg/mL) for 48 h, the cell cycle of Jurkat, SU-DHL-4 and Raji cells was arrested in G2/M phase(all P<0.01). The migration and invasion activities of Jurkat and Raji cells were significantly suppressed by Qi-Zhen-Gui-Pi decoction(15 mg/mL)(all P<0.01). According to the gene expression profiling data, 648 genes were up-regulated, and 1

关 键 词：淋巴瘤 植物药疗法 细胞增殖 细胞凋亡 细胞运动 

分 类 号：R733.4[医药卫生—肿瘤]