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徐长卿中的天然产物 XCQ-9 对 Jurkat 细胞增殖和凋亡的影响及
机制研究
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韦学耐 1, 2* ,杨 坤 ,刘 琴 ,赵 鹏 ,晏 英 ,李艳梅 (1.贵州医科大学药学院,贵阳 550025;2.贵州
省中国科学院天然产物化学重点实验室,贵阳 550014;3.贵州医科大学医药卫生管理学院,贵阳 550025)
中图分类号 R965 文献标志码 A 文章编号 1001-0408(2023)01-0047-05
DOI 10.6039/j.issn.1001-0408.2023.01.09
摘 要 目的 探讨徐长卿中天然产物XCQ-9抑制人急性T淋巴细胞白血病Jurkat细胞增殖和凋亡的作用及可能机制。方法 以
Jurkat细胞作为白血病细胞模型,采用MTT法测定0(空白对照)、2.5、5、10、20、40 μmol/L XCQ-9作用24、48、72 h后对Jurkat细胞
增殖的抑制作用。用0(空白对照)、2.5、5、10 μmol/L XCQ-9作用于Jurkat细胞24、48 h后,利用流式细胞术分析XCQ-9对细胞周
期和细胞凋亡的影响,并通过 Western blot 实验检测上述药物作用 24 h 后细胞中胱天蛋白酶 9(Caspase-9)、活化的 Caspase-9
(Cleaved Caspase-9)、Caspase-3、活化的Caspase-3(Cleaved Caspase-3)、聚腺苷二磷酸-核糖聚合酶(PARP)、活化的PARP(Cleaved
PARP)、细胞周期蛋白依赖性激酶1(CDK1)和细胞周期蛋白B1(Cyclin B1)的表达情况。结果 与空白对照比较,不同浓度XCQ-9
均可显著降低Jurkat细胞的存活率(P<0.01),并呈时间和浓度依赖性趋势。5、10 μmol/L XCQ-9作用48 h后均可显著诱导Jurkat细胞
凋亡(P<0.05或P<0.01),将细胞周期阻滞在G2期(P<0.01)。10 μmol/L XCQ-9作用24 h后,可显著下调细胞中CDK1、Caspase-9
蛋白的表达(P<0.01),上调细胞中 Cyclin B1、Cleaved Caspase-9、Cleaved Caspase-3 和 Cleaved PARP 蛋白的表达(P<0.05 或 P<
0.01)。结论 XCQ-9通过诱导G2期阻滞抑制Jurkat细胞增殖,并激活Caspase通路促进细胞凋亡,从而发挥其抗肿瘤作用。
关键词 XCQ-9;徐长卿;人急性T淋巴细胞白血病;Jurkat细胞;细胞凋亡;细胞周期;胱天蛋白酶途径
Study on the effect of natural compound XCQ-9 of Cynanchum paniculatum on the proliferation and
apoptosis of Jurkat cell and its mechanism
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WEI Xuenai ,YANG Kun ,LIU Qin ,ZHAO Peng ,YAN Ying ,LI Yanmei (1. School of Pharmaceutical,
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Guizhou Medical University, Guiyang 550025, China;2. Key Laboratory of Chemistry for Natural Products,
Guizhou Province and Chinese Academy of Sciences, Guiyang 550014, China;3. School of Medicine and
Health Management, Guizhou Medical University, Guiyang 550025, China)
ABSTRACT OBJECTIVE To investigate the inhibitory effect of natural compound XCQ-9 of Cynanchum paniculatum on the
proliferation and apoptosis of Jurkat cell line of human T-cell acute lymphoblastic leukemia and its possible mechanism.
METHODS Jurkat cell was used as the leukemia cell model, and MTT assay was adopted to detect the inhibitory effects of 0
(blank control), 2.5, 5, 10, 20 and 40 μmol/L XCQ-9 on the proliferation of Jurkat cell after treated for 24, 48, 72 h. After
treated with 0 (blank control), 2.5, 5, 10 μmol/L XCQ-9 for 24 h and 48 h, the cell cycle and apoptosis were analyzed by flow
cytometry. The expressions of Caspase-9, Cleaved Caspase-9, Caspase-3, Cleaved Caspase-3, poly ADP-ribose poly-merase
(PARP), Cleaved-PARP, cyclin-dependent kinase 1 (CDK1) and Cyclin B1 were detected by Western blot after treated for 24 h.
RESULTS Compared with blank control group, XCQ-9 at different concentrations could significantly decrease the survival rate of
Jurkat cells (P<0.01), and showed a dose and time-dependent manner. After 48 h treatment of 5, 10 μmol/L XCQ-9, Jurkat cell
apoptosis was induced significantly (P<0.05 or P<0.01), and the cell was arrested in G2 phase (P<0.01). After 24 h treatment of
10 μmol/L XCQ-9, the protein expressions of CDK1 and Caspase-9 were remarkably down-regulated (P<0.01), while the protein
expressions of Cyclin B1, Cleaved Caspase-9, Cleaved Caspase-3 and Cleaved PARP were significantly up-regulated (P<0.05 or
P<0.01). CONCLUSIONS XCQ-9 plays anti-tumor effect
Δ 基金项目 国 家 自 然 科 学 基 金 资 助 项 目(No. 81872772,No.
through inducing G2 phase arrest to inhibit proliferation and
81960546,No.U1812403);贵州省科技计划项目(No.QKHPTRC〔2020〕
5008) activating Caspase pathway to increase apoptosis.
*第一作者 硕士研究生。研究方向:小分子化合物抗肿瘤基础。 KEYWORDS XCQ-9; Cynanchum paniculatum; human T-
E-mail:2531392591@qq.com cell acute lymphoblastic leukemia; Jurkat cell; cell apoptosis;
# 通信作者 研究员,硕士生导师,博士。研究方向:小分子化合物 cell cycle; Caspase pathway
抗肿瘤基础。电话:0851-83834026。E-mail:liyanmei518@hotmail.
com
中国药房 2023年第34卷第1期 China Pharmacy 2023 Vol. 34 No. 1 · 47 ·