干扰TGF-βⅡ型受体表达对NB4细胞增殖、分化及凋亡的影响  

Effect of interfering TGF-β receptor Ⅱ expression on viability,differentiation and apoptosis of NB4 cells

作　　者：陈美玲 张淑遐 郭雅斐[1] 陈莹莹[1] 吴勇[1] CHEN Mei-ling;ZHANG Shu-xia;GUO Ya-fei;CHEN Ying-ying;WU Yong(Fujian Institute of Hematology,Fujian Provincial Key Laboratory on Hematology,Fujian Medical University Union Hospital,Fuzhou 350004,China.)

机构地区：[1]福建省血液病研究所,福建省血液病学重点实验室,福建医科大学附属协和医院,福建福州350004

出　　处：《中国病理生理杂志》2018年第10期1729-1735,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81270571);福建省血液医学中心建设项目资助[闽政办(2017)4号];国家临床重点专科建设资助项目[财社(2011)170号];福建省临床重点专科建设资助项目[闽卫科教(2012)149号]

摘　　要：目的:研究干扰TGF-βⅡ型受体(TβRⅡ)表达对人急性早幼粒细胞白血病NB4细胞生长、分化及凋亡的影响。方法:应用慢病毒介导RNA干扰技术,下调NB4细胞TβRⅡ基因的表达,获得TβRⅡ-shRNA NB4细胞,CCK-8法检测细胞的活力,流式细胞术检测CD11b表达,并用瑞氏-吉姆萨染色法检测全反式维甲酸对细胞分化的影响; Annexin V-FITC/PI双荧光标记法及AO/EB染色观察三氧化二砷对细胞凋亡的影响。结果:TβRⅡ-shRNA NB4细胞的活力高于NB4细胞。采用不同浓度(0. 01、0. 02、0. 04、0. 08和0. 1μmol/L)的全反式维甲酸进行96 h的孵育后,2组细胞均出现分化现象(CD11b表达增加),并呈剂量依赖性,但TβRⅡ-shRNA NB4细胞的分化率低于NB4细胞。不同浓度(2、4和8μmol/L)的三氧化二砷孵育24 h后,2组细胞均出现凋亡现象(AO/EB染色),呈剂量依赖性,但TβRⅡ-shRNA NB4细胞凋亡率显著低于NB4细胞,其中用8μmol/L三氧化二砷孵育TβRⅡ-shRNA NB4细胞和NB4细胞24 h,凋亡率分别是(49. 15±2. 05)%和(66. 85±2. 41)%(P <0. 01)。结论:下调TβRII可以促进NB4细胞的生长,部分拮抗全反式维甲酸诱导的细胞分化及三氧化二砷诱导的细胞凋亡。AIM:To evaluate the effect of interfering TGF-βreceptorⅡ(TβRII)expression on the viability and differentiation of human acute promyelocytic leukemia NB4 cells induced by all-trans retinoic acid(ATRA)and their apoptosis induced by arsenic trioxide(ATO).METHODS:The technique of lentivirus-mediated RNA interference was used to obtain stable NB4 cells with TβRII knockdown,named TβRII-shRNA NB4 cells.CCK-8 assay was used to detect the viability of TβRII-shRNA NB4 cells.The expression level of CD11b was analyzed by flow cytometry,and Wright-Giemsa staining was used to detect the effects of ATRA on the differentiation of TβRII-shRNA NB4 cells.Double staining(Annexin V-FITC/PI)and AO/EB staining were used to detect the effects of ATO on the apoptosis of TβRII-shRNA NB4 cells.RESULTS:The viability of TβRII-shRNA NB4 cells was significantly higher than that of NB4 parental cells.The differentiation was induced in TβRII-shRNA NB4 cells and NB4 parent cells by treatment with ATRA at different concentration(0.01,0.02,0.04,0.08,0.1μmol/L)for 96 h.The differentiation rate of TβRII-shRNA NB4 cells was lower than that of NB4 parental cells in a dose-dependent manner.ATO induced apoptosis of TβRII-shRNA NB4 cells and NB4 parent cells at different concentrations(2,4 and 8μmol/L)for 24 h.The apoptotic rate of TβRII-shRNA NB4 cells was lower than that of NB4 parental cells dose-dependently.At the concentration of 8μmol/L for 24 h,the apoptotic rates in TβRII-shRNA NB4 cells and NB4 cells were(49.15±2.05)%and(66.85±2.41)%,respectively(P<0.01).CONCLUSION:Down-regulation of TβRII increases the viability of NB4 cells,inhibits NB4 cell differentiation induced by ATRA,and also inhibits apoptosis induced by ATO.

关 键 词：转化生长因子Β受体 NB4细胞 全反式维甲酸 分化 细胞凋亡 

分 类 号：R577.3[医药卫生—消化系统] R363[医药卫生—内科学]