中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (31): 6674-6680.doi: 10.12307/2025.661

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

盐酸青藤碱抑制急性T淋巴细胞白血病CEM细胞株的作用及转录组学分析

康林之1,刘振帅1,魏佳旭1,常  娜2,朱大诚1   

  1. 1江西中医药大学,江西省南昌市  330004;2延安大学,陕西省延安市  716000
  • 收稿日期:2024-05-28 接受日期:2024-08-12 出版日期:2025-11-08 发布日期:2025-02-24
  • 通讯作者: 朱大诚,教授,博士生导师,江西中医药大学,江西省南昌市 330004
  • 作者简介:康林之,女,1979年生,陕西省岐山县人,汉族,江西中医药大学在读博士,讲师,主要从事中医药抗肿瘤研究。
  • 基金资助:
    江西省“双一流”学科(中医学)建设资助项目(JXYLXK-ZHYI051),项目参与人:朱大诚;2022江西中医药大学校级研究生创新专项资金项目(JZYC22B01),项目负责人:康林之

Inhibitory effects of sinomenine hydrochloride in T-cell acute lymphoblastic leukemia CEM cells and transcriptomic analysis

Kang Linzhi1, Liu Zhenshuai1, Wei Jiaxu1, Chang Na2, Zhu Dacheng1   

  1. 1Jiangxi University of Chinese Medicine, Nanchang 330004, Jiangxi Province, China; 2Yan'an University, Yan'an 716000, Shanxi Province, China
  • Received:2024-05-28 Accepted:2024-08-12 Online:2025-11-08 Published:2025-02-24
  • Contact: Zhu Dacheng, Professor, Doctoral supervisor, Jiangxi University of Chinese Medicine, Nanchang 330004, Jiangxi Province, China
  • About author:Kang Linzhi, Doctoral candidate, Lecturer, Jiangxi University of Chinese Medicine, Nanchang 330004, Jiangxi Province, China
  • Supported by:
    Jiangxi Province “Double First-Class” Discipline (Traditional Chinese Medicine) Construction Project, No. JXYLXK-ZHYI051 (to ZDC); Jiangxi University of Chinese Medicine Postgraduate Innovation Special Fund Project, No. JZYC22B01 (to KLZ)

摘要:

文题释义:

盐酸青藤碱:是从中药青风藤中提取的有效生物碱单体的盐酸盐。药理研究表明盐酸青藤碱具有祛风除湿、活血通络、利尿消肿以及抑制肿瘤的作用。
转录组学分析:通过高通量测序分析细胞在某一功能状态下的转录本序列信息,如转录本的结构和表达水平,揭示特定的生物学过程,广泛应用于基础研究、临床诊断和药物研发等领域。

摘要
背景:盐酸青藤碱有抗多种肿瘤的作用,但目前尚不清楚盐酸青藤碱对急性T淋巴细胞白血病的作用。
目的:探讨盐酸青藤碱对急性T淋巴细胞白血病CEM细胞的抑制作用。
方法:应用不同浓度(0.5,1,2,4 mmol/L)盐酸青藤碱处理CEM细胞,CCK-8检测细胞增殖抑制率并计算IC50;倒置显微镜和吉姆萨染色观察CEM细胞形态变化;利用RNA-seq测序分析差异基因表达并进行生物信息学分析。结合转录组测序结果,流式细胞术检测不同浓度(1,2,4 mmol/L)盐酸青藤碱作用后CEM细胞凋亡率;Western blot检测不同浓度(1,2,4 mmol/L)盐酸青藤碱作用后CEM细胞中Bcl-2、Bax、Caspase-9蛋白的表达。
结果与结论:①盐酸青藤碱呈剂量和时间依赖性地抑制CEM细胞生长;②盐酸青藤碱干预后CEM细胞数量下降,核固缩;③RNA-seq测序筛出53个异常表达基因,基因本体分析主要富集在细胞过程、细胞解剖实体与粘连关系等,信号通路分析与肿瘤相关的是细胞凋亡;④盐酸青藤碱呈剂量依赖性地促进CEM细胞凋亡;⑤盐酸青藤碱上调CEM细胞中Bax、Caspase-9蛋白表达,下调Bcl-2蛋白表达。由此可见,盐酸青藤碱可诱导CEM细胞凋亡从而抑制细胞增殖,可能与其上调Bax和Caspase-9蛋白表达,以及下调 Bcl-2 蛋白表达有关。

关键词: 盐酸青藤碱, 急性T淋巴细胞白血病, CEM细胞, 细胞凋亡, RNA-seq测序, 增殖抑制, 工程化细胞

Abstract: BACKGROUND: Sinomenine hydrochloride has anti-tumor effects, but it is rarely reported in T-cell acute lymphoblastic leukemia. 
OBJECTIVE: To investigate the inhibitory effect of sinomenine hydrochloride on CEM cells in acute T lymphoblastic leukemia. 
METHODS: Different concentrations (0.5, 1, 2 and 4 mmol/L) of sinomenine hydrochloride were used to act on CEM cells. CCK-8 assay was used to detect the inhibition rate of cell proliferation and calculate the IC50. Inverted microscope and Giemsa staining were used to observe the changes of CEM cells. The RNA sequencing was performed to analyze the differential gene expression and biological information. Combined with transcriptome sequencing analysis results, flow cytometry was used to detect the apoptosis rate of CEM cells after treatment with of different concentrations (1, 2, and 4 mmol/L) of sinomenine hydrochloride. Western blot assay was used to detect the expression of Bcl-2, Bax, and Caspase-9 proteins in CEM cells after treatment with of different concentrations (1, 2, and 4 mmol/L) of sinomenine hydrochloride.  
RESULTS AND CONCLUSION: (1) Sinomenine hydrochloride inhibited the growth of CEM cells in dose and time-dependent manner. (2) After dosing, the number of CEM cells decreased and pyknosis appeared. (3) RNA sequencing revealed 53 differential expressed genes. Gene Ontology was significantly enriched in cellular process, cellular anatomical entities, and binding. Signaling pathway analysis related to tumor was apoptosis. (4) Sinomenine hydrochloride induced the apoptosis of CEM cell in a concentration-dependent manner. (5) Sinomenine hydrochloride could promote the expressions of Bax and Caspase-9, but restrain the expression of Bcl-2 in CEM cells. Therefore, sinomenine hydrochloride can induce apoptosis in CEM cells and suppress cell proliferation, maybe via up-regulation of the protein levels of Bax and caspase-9 and down-regulation of the protein level of Bcl-2. 


Key words: sinomenine hydrochloride, T-cell acute lymphoblastic leukemia, CEM cell, apoptosis, RNA sequencing, proliferation inhibition, engineered cell

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