漆姑草醇提物对人急性早幼粒白血病细胞的诱导分化作用

doi:10.3969/j.issn.1004-616x.2021.01.009

癌变·畸变·突变 ›› 2021, Vol. 33 ›› Issue (1): 42-47.doi: 10.3969/j.issn.1004-616x.2021.01.009

漆姑草醇提物对人急性早幼粒白血病细胞的诱导分化作用

唐文娟¹, 伏琼¹, 程勇¹, 罗世成¹, 梁冰^1,2

1. 贵州医科大学药理学教研室, 贵州贵阳 550025;
2. 贵州医科大学环境污染与疾病监控教育部重点实验室, 贵州贵阳 550025

收稿日期:2020-11-10 修回日期:2020-12-02 出版日期:2021-01-30 发布日期:2021-02-06
通讯作者: 梁冰,E-mail:bliang163@163.com E-mail:bliang163@163.com
作者简介:唐文娟,E-mail:413701071@qq.com。
基金资助:
贵阳市科技计划项目（筑科合同[2017]30-9号）；贵州省科技创新人才团队项目[黔科合平台人才（2016）5625号]；贵州省2017年度“千”层次创新型人才项目

Induction of differentiation of human acute promyelocytic leukemia cells by an ethanol extract from Herba Saginae Japonicae

TANG Wenjuan¹, FU Qiong¹, CHENG Yong¹, LUO Shicheng¹, LIANG Bing^1,2

1. Department of Pharmacology, Guizhou Medical University, Guiyang 550025;
2. Key Laboratory of Environmental Pollution and Disease Monitoring, Ministry of Education; Guizhou Medical University, Guiyang 550025, Guizhou, China

Received:2020-11-10 Revised:2020-12-02 Online:2021-01-30 Published:2021-02-06

摘要/Abstract

摘要： 目的： 探讨漆姑草醇提物（HSJ）对人急性早幼粒白血病细胞（NB4）的诱导分化作用。方法： 以NB4细胞为研究对象，设HSJ低（30 μg/mL）、中（60 μg/mL）、高（120 μg/mL）剂量组和阴性对照组，共4组；HSJ作用NB4细胞48 h后，采用透射电镜法观察NB4细胞形态；硝基四唑氮（NBT）还原实验检测NB4细胞分化能力；流式细胞术检测NB4细胞周期分布；流式细胞术检测NB4细胞表面分化抗原CD14和CD11b细胞的表达率；实时荧光定量PCR （qPCR）检测c-fos mRNA表达；Western blot检测c-fos蛋白表达。结果： 与对照组比较，3个不同剂量HSJ处理组NB4细胞胞核均缩小，胞浆呈空泡状，核形呈肾形或蚕豆形；各HSJ处理组NB4细胞的NBT还原率均高于对照组，差异有统计学意义（P < 0.05）；HSJ处理组NB4细胞中G2期细胞比例均高于对照组，而S期细胞比例均低于对照组，差异有统计学意义（P < 0.05）；HSJ处理组CD14和CD11b阳性细胞表达率均高于对照组（P < 0.05）；HSJ低剂量组cfos mRNA和蛋白表达与对照组比较，差异均无统计学意义（P > 0.05），HSJ中、高剂量组c-fos mRNA和蛋白表达均高于对照组，差异均有统计学意义（P < 0.05）。结论： HSJ可能诱导NB4细胞向成熟粒细胞方向分化。

关键词: 漆姑草, 流式细胞术, NB4细胞, 诱导分化, 急性早幼粒白血病

Abstract: OBJECTIVE: To study cellular changes during induction of differentiation of human acute promyelocytic leukemia cells (NB4) by an alcohol extract from Herba Saginae Japonicae (HSJ). METHODS: NB4 cells in culture were were organized into four groups:HSJ low dose (30 μg/mL), HSJ middle dose (60 μg/mL), HSJ high dose (120 μg/mL) and negative control groups. Cells in the experimental groups were treated with HSJ for 48 h and their cellular morphology was observed by transmission electron microscopy. nitrotriazole nitrogen (NBT) reduction test was used to detect the differentiation ability of NB4 cells. The distribution of NB4 cell cycles and expression rates of CD14 and CD11b cells were detected using flow cytometry. Expression of c-fos mRNA was detected using quantitative real-time PCR(qPCR). Expression of cfos protein was detected using the Western blot method. RESULTS: Compared with the control group, the nuclei of HSJ-treated cells, in the three different doses, were reduced and changed into kidney-shape, and the cytoplasm was vacuolated. The NBT test showed that the reduction rate of the treated cells were significantly higher than that in the control group (P < 0.05). The proportions of G2 phase cells in the treated groups were significantly higher than that in the control group, while the proportion of S phase cells were significantly lower (P < 0.05). The expression rates of CD14 and CD11b positive cells in the treated groups were significantly higher than that in the control group (P < 0.05). In addition, expression rates of c-fos mRNA and c-fos proteins in the middleand the high-dose groups were significantly higher than that in the control group (P < 0.05) but no difference for the low-dose group. CONCLUSION: HSJ may induce NB4 cells to differentiate into mature granulocytes.

Key words: Herba Saginae Japonicae, flow cytometry, NB4 cells, differentiation, acute promyelocytic leukemia

中图分类号:

R73-3

唐文娟, 伏琼, 程勇, 罗世成, 梁冰. 漆姑草醇提物对人急性早幼粒白血病细胞的诱导分化作用[J]. 癌变·畸变·突变, 2021, 33(1): 42-47.

TANG Wenjuan, FU Qiong, CHENG Yong, LUO Shicheng, LIANG Bing. Induction of differentiation of human acute promyelocytic leukemia cells by an ethanol extract from Herba Saginae Japonicae[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(1): 42-47.

参考文献

[1] ORFALI N, O'DONOVAN T R, NYHAN M J, et al. Induction of autophagy is a key component of all-trans-retinoic acidinduced differentiation in leukemia cells and a potential target for pharmacologic modulation[J]. Exp Hematol, 2015, 43(9):781-793.
[2] NITTO T, SAWAKI K. Molecular mechanisms of the antileukemia activities of retinoid and arsenic[J]. J Pharmacol Sci, 2014, 126(3):179-185.
[3] 吴晓丹. 全反式维甲酸联合三氧化二砷治疗初发急性早幼粒细胞白血病的近期疗效分析[J]. 中国医药指南, 2018, 16(1):96-97.
[4] 陈冬, 陆滢, 裴仁治, 等. 小剂量高三尖杉酯碱联合全反式维甲酸及三氧化二砷治疗急性早幼粒细胞白血病的临床研究[J]. 中国临床药理学与治疗学, 2018, 23(5):561-569.
[5] 姚仕菲, 刘北忠, 陈敏, 等. EGCG通过PTEN增强ATRA对早幼粒白血病细胞株的分化作用[J]. 基因组学与应用生物学, 2018, 37(5):2182-2186.
[6] LO-COCO F, AVVISATI G, VIGNETTI M, et al. Retinoic acid and arsenic trioxide for acute promyelocytic leukemia[J]. N Engl J Med, 2013, 369(2):111-121.
[7] 宋梅, 吴秀梅, 张梅, 等. 白血病细胞早幼粒细胞白血病(PML)蛋白存在形式的检测和临床意义[J]. 细胞与分子免疫学杂志, 2017, 33(7):971-976.
[8] 伏琼, 唐文娟, 程勇, 等. 漆姑草醇提物对HL-60细胞诱导分化作用[J]. 中国公共卫生, 2017, 33(7):1066-1069.
[9] 王雷鸣, 梁冰, 李淑芳, 等. 漆姑草水提物对K562细胞凋亡及细胞周期影响[J]. 中国公共卫生, 2014, 30(12):1520-1522.
[10] 于丽, 吴静, 王迪, 等. 急性早幼粒细胞白血病细胞分化的调控机制新进展[J]. 中国免疫学杂志, 2019, 35(8):1003-1006.
[11] 赵钰, 林玮. 白血病细胞的定向诱导分化及PADI4在诱导分化中的作用[J]. 中南医学科学杂志, 2019, 47(1):85-88.
[12] 范奎, 代良敏, 伍振峰, 等. 放化疗所致骨髓抑制的研究进展[J]. 中华中医药杂志, 2017, 32(1):210-214.
[13] 魏婧昕, 丁兰, 刘扬, 等. Rabdosin B对急性早幼粒白血病细胞HL-60分化的诱导作用[J]. 中草药, 2018, 49(11):25912600.
[14] 陈智, 王博, 裘玫, 等. 对香豆酸组合物对白血病细胞的增殖抑制作用及机制[J]. 中国药学杂志, 2017, 52(17):1503-1509.
[15] 申振铭, 庄步玺, 赵瑶, 等. 复方中药制剂对急性髓系白血病细胞诱导分化与增殖的作用[J]. 中国实验血液学杂志, 2019, 27(2):403-408.
[16] 高志强, 程会云, 李同源, 等. 知母皂苷AⅢ对白血病细胞HL-60增殖、凋亡和细胞周期的影响及机制探讨[J]. 中成药, 2017, 39(7):1487-1490.
[17] IRIYAMA N, YUAN B, HATTA Y, et al. Lyn, a tyrosine kinase closely linked to the differentiation status of primary acute myeloid leukemia blasts, associates with negative regulation of all-trans retinoic acid (ATRA) and dihydroxyvitamin D3(VD3)-induced HL-60 cells differentiation[J]. Cancer Cell Int, 2016, 16:37.
[18] 余潇苓, 赵燕娜, 郑智茵, 等. 地西他滨抑制K562白血病细胞增殖和诱导分化的作用[J]. 中国病理生理杂志, 2017, 33(1):13-17.
[19] HEO S K, NOH E K, YOON D J, et al. Rosmarinic acid potentiates ATRA-induced macrophage differentiation in acute promyelocytic leukemia NB4 cells[J]. Eur J Pharmacol, 2015, 747:36-44.
[20] 郑卓军, 顾伟英, 谢晓宝, 等. Erbin基因对急性髓系白血病细胞周期、增殖、凋亡和分化的影响[J]. 中华实验外科杂志, 2018, 35(2):319-322.
[21] BLAIR O C, CARBONE R, SARTORELLI A C. Differentiation of HL-60 promyelocytic leukemia cells monitored by flow cytometric measurement of nitro blue tetrazolium (NBT) reduction[J]. Cytometry, 1985, 6(1):54-61.
[22] STUDZINSKI G P, HARRISON L E. Differentiationrelated changes in the cell cycle traverse[J]. Int Rev Cytol, 1999, 189:1-58.
[23] STOICA S, MAGOULAS G E, ANTONIOU A I, et al. Synthesis of minoxidil conjugates and their evaluation as HL-60 differentiation agents[J]. Bioorg Med Chem Lett, 2016, 26(4):1145-1150.
[24] 邓守恒, 俞远东, 张军, 等. 硒化壳聚糖对急性早幼粒细胞白血病NB4细胞体外分化的影响[J]. 山西医药杂志, 2010, 39(19):911-913.

漆姑草醇提物对人急性早幼粒白血病细胞的诱导分化作用

Induction of differentiation of human acute promyelocytic leukemia cells by an ethanol extract from Herba Saginae Japonicae

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	伏琼, 唐文娟, 程勇, 贺仁政, 梁冰. 漆姑草醇提物对白血病细胞K562的诱导分化作用[J]. 癌变·畸变·突变, 2021, 33(1): 37-41.
[2]	黄鹏程, 周长慧, 李申宁, 常艳. 基于流式细胞术的组蛋白γ-H2AX磷酸化检测方法的建立[J]. 癌变·畸变·突变, 2017, 29(4): 284-288.
[3]	杨英杰, 刘建香, 高刚. 电离辐射对小鼠外周血单核细胞亚群的影响[J]. 癌变·畸变·突变, 2016, 28(4): 298-301.
[4]	洪丽玲, 王婷, 范宾, 周庆云, 黄永焯, 王峰. 二溴乙腈对L5178Y细胞的遗传毒性及促凋亡作用[J]. 癌变·畸变·突变, 2015, 27(6): 450-453,458.
[5]	欧红梅，周长慧，涂宏刚，黄鹏程，常艳. 流式细胞术检测体外微核方法的建立[J]. 癌变·畸变·突变, 2015, 27(1): 39-43.
[6]	邓婷婷，谢妮，黄艳华，黄海燕，李子刚，胡章立，袁建辉. 毛细管电泳法检测耐米托蒽醌乳腺癌细胞系全基因组甲基化水平[J]. 癌变·畸变·突变, 2014, 26(2): 108-112.
[7]	张铭,周长慧,常艳,王征,涂宏刚,李婕. 体内Pig-a基因突变试验高通量方法联合验证研究[J]. 癌变·畸变·突变, 2013, 25(5): 392-395.
[8]	王雷,王治东,胡海亮,张学清,陈茂胜,薛绍礼,陈英. 外周血网织红细胞和T淋巴细胞亚群用于辐射损伤快速剂量估算的可行性研究[J]. , 2011, 23(6): 452-455.
[9]	李超霞;方　力;杨小红. MDM2、P53和P27蛋白在食管鳞癌及其癌旁组织中的表达及其意义[J]. , 2009, 21(5): 362-366.
[10]	段玉清1,2/ 张海晖1/ 吴　佳2/ 谢笔钧2. 莲房原花青素对黑色素瘤B16细胞的诱导分化作用[J]. , 2007, 19(5): 377-380.
[11]	江城梅1;赵文红1;庄　颖1;吕合作2. 三羟异黄酮对黑色素瘤B16细胞的诱导分化作用[J]. , 2006, 18(1): 46-049.
[12]	杜　芸;王小玲;王永军;吴国祥;杨会钗;左连富. 流式细胞术测定COX-2,iNOS基因蛋白在食管癌上的表达及其与Bcl-2,PCNA的相关性[J]. , 2005, 17(4): 224-226.
[13]	贾庆军，杨录军，刘晋，周燕虹，刘胜学，周紫桓，曹　佳. 硝酸羟胺对雄性小鼠生殖细胞和精子影响的初步研究[J]. , 2004, 16(6): 362-364.
[14]	姚　莉;卞修武;张树辉;陈自强;梁晓俐. 应用AP-PCR法分析去甲二氢愈创木酸对胶质瘤细胞基因组甲基化状态的影响[J]. , 2004, 16(2): 81-82.
[15]	艾　军, 何兰欣, 梁索源, 任金荣. 抗癌药物对细胞株增殖的影响及凋亡诱导作用[J]. , 2002, 14(3): 200-201.