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  

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

作　　者：康林之[1] 刘振帅 魏佳旭 常娜 朱大诚[1] Kang Linzhi;Liu Zhenshuai;Wei Jiaxu;Chang Na;Zhu Dacheng(Jiangxi University of Chinese Medicine,Nanchang 330004,Jiangxi Province,China;Yan'an University,Yan'an 716000,Shanxi Province,China)

机构地区：[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

出　　处：《中国组织工程研究》2025年第31期6674-6680,共7页Chinese Journal of Tissue Engineering Research

基　　金：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。

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。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.

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 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R551