ACT001 inhibits the neurosphere formation and stemness maintenance of U87-MG glioma stem cells through STAT3 signaling pathway

Q4 Medicine 中华神经外科杂志 Pub Date : 2019-11-28 DOI:10.3760/CMA.J.ISSN.1001-2346.2019.11.019

Jiabo Li, Luqing Tong, L. Yi, Peidong Liu, Yang Xie, Xuya Wang

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Abstract

Objective To investigate the effect of sesquiterpene lactone derivative (ACT001) on the neurosphere formation and stemness maintenance of U87-MG glioma stem cells. Methods Bioinformatics was used to analyze the relationship between STAT3 mRNA and different grades of human glioma as well as the overall survival in 1 477 glioma patients in CGGA, TCGA, and GEO (GSE4290, GSE16011) databases. Magnetic activated cell sorting method was used to obtain CD133-positive U87-MG cells. The effect of ACT001 on the survival viability of U87-MG GSCs was detected via Cell Counting Kit-8 (CCK-8). The effect of ACT001 on the ability of neurosphere formation was observed by single-cell neurosphere formation assay. Immunofluorescence staining was used to measure the effect of ACT001 on stem cells markers (CD133、CD44 and Nestin) in neurospheres of U87-MG GSCs. The expressions of STAT3, p-STAT3, CD44 and CD133 were detected using western blot after treatment of ACT001. Results Bioinformatics analysis showed that as the grades of glioma increased, the expression of STAT3 mRNA also increased (all P 0.05), while the protein expression of p-STAT3 was decreased through assessment by Western blot (all P<0.01). Conclusion ACT001 could suppress the neurosphere formation ability and reduce the stemness maintenance in U87-MG glioma stem cells by inhibiting STAT3 signaling pathway. Key words: Glioma; Neoplastic stem cells; Sesquiterpene lactone derivative; STAT3 signaling pathway

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ACT001通过STAT3信号通路抑制U87-MG胶质瘤干细胞的神经球形成和干性维持

目的探讨倍半萜内酯衍生物(ACT001)对U87-MG胶质瘤干细胞神经球形成和干细胞维持的影响。方法采用生物信息学方法，分析CGGA、TCGA和GEO (GSE4290、GSE16011)数据库中1 477例胶质瘤患者的STAT3 mRNA与胶质瘤不同分级及总生存率的关系。采用磁活化细胞分选法获得cd133阳性U87-MG细胞。通过细胞计数试剂盒-8 (CCK-8)检测ACT001对U87-MG GSCs存活能力的影响。通过单细胞神经球形成实验观察ACT001对大鼠神经球形成能力的影响。采用免疫荧光染色法检测ACT001对U87-MG GSCs神经球干细胞标志物CD133、CD44和Nestin的影响。western blot检测细胞中STAT3、p-STAT3、CD44、CD133的表达。结果生物信息学分析显示，随着胶质瘤分级的增加，STAT3 mRNA的表达也增加(均P< 0.05)，而Western blot评估P -STAT3蛋白的表达降低(均P<0.01)。结论ACT001可通过抑制STAT3信号通路抑制U87-MG胶质瘤干细胞的神经球形成能力，降低干细胞的维持能力。关键词:胶质瘤;肿瘤干细胞;倍半萜内酯衍生物;STAT3信号通路

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来源期刊

中华神经外科杂志 Medicine-Surgery

CiteScore

0.10

自引率

0.00%

发文量

10706

期刊介绍： Chinese Journal of Neurosurgery is one of the series of journals organized by the Chinese Medical Association under the supervision of the China Association for Science and Technology. The journal is aimed at neurosurgeons and related researchers, and reports on the leading scientific research results and clinical experience in the field of neurosurgery, as well as the basic theoretical research closely related to neurosurgery.Chinese Journal of Neurosurgery has been included in many famous domestic search organizations, such as China Knowledge Resources Database, China Biomedical Journal Citation Database, Chinese Biomedical Journal Literature Database, China Science Citation Database, China Biomedical Literature Database, China Science and Technology Paper Citation Statistical Analysis Database, and China Science and Technology Journal Full Text Database, Wanfang Data Database of Medical Journals, etc.