Department of Pediatrics (Children Hematological Oncology), Birth Defects and Childhood Hematological Oncology Laboratory, The Affiliated Hospital of Southwest Medical University, Sichuan Clinical Research Center for Birth Defects, Luzhou, Sichuan, China.
State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China.
Pharm Biol. 2023 Dec;61(1):259-270. doi: 10.1080/13880209.2023.2168703.
Due to the poor prognosis of T-cell acute lymphoblastic leukaemia (T-ALL), there is an urgent need to identify safer and more cost-effective drugs.
This study evaluated the antitumour activity of Shuanghuanglian (SHL) on T-ALL cells and elucidated the mechanism.
Jurkat and Molt4 cells were treated with SHL (0.1, 0.2 and 0.4 mg/mL) for 24 and 48 h. The controls were treated with RPMI 1640 containing 10% foetal bovine serum. Cell viability was evaluated through Cell Counting Kit-8 assay. Patterns of death and signalling pathway alterations caused by SHL were identified by network pharmacology combined with GO enrichment analysis and then were verified by Hoechst 33342 staining, Annexin V-FITC/PI staining and Western blotting. Interactions of the active ingredients with targets were analysed by molecular docking.
The IC values of SHL in Jurkat and Molt4 cells were 0.30 ± 0.10 and 0.48 ± 0.07 mg/mL, respectively, at 24 h and 0.27 ± 0.05 and 0.30 ± 0.03 mg/mL at 48 h. In T-ALL, 117 target genes of SHL were mainly enriched in the apoptosis and NOTCH signalling pathways. SHL induced apoptosis was confirmed by Hoechst 33342 staining and flow cytometry. The protein levels of cleaved caspase-7 and cleaved PARP were significantly increased but those of cleaved NOTCH1 and MYC were reduced. The active ingredients of SHL can interact with γ-secretase. SHL induces apoptosis in T-ALL cells the NOTCH1-MYC pathway and may be a potential drug for the treatment of T-ALL.
由于 T 细胞急性淋巴细胞白血病（T-ALL）的预后较差，因此迫切需要确定更安全、更具成本效益的药物。
本研究评估了双黄连（SHL）对 T-ALL 细胞的抗肿瘤活性，并阐明了其作用机制。
Jurkat 和 Molt4 细胞分别用 SHL（0.1、0.2 和 0.4mg/mL）处理 24 和 48 小时。对照组用含 10%胎牛血清的 RPMI 1640 处理。通过细胞计数试剂盒-8 测定法评估细胞活力。通过网络药理学结合 GO 富集分析，鉴定 SHL 引起的死亡模式和信号通路改变，并通过 Hoechst 33342 染色、Annexin V-FITC/PI 染色和 Western blot 进行验证。通过分子对接分析活性成分与靶标的相互作用。
SHL 在 Jurkat 和 Molt4 细胞中的 IC 值在 24 小时分别为 0.30±0.10 和 0.48±0.07mg/mL，在 48 小时分别为 0.27±0.05 和 0.30±0.03mg/mL。在 T-ALL 中，SHL 的 117 个靶基因主要富集在细胞凋亡和 NOTCH 信号通路中。Hoechst 33342 染色和流式细胞术证实 SHL 诱导细胞凋亡。Cleaved caspase-7 和 cleaved PARP 的蛋白水平显著升高，但 cleaved NOTCH1 和 MYC 的蛋白水平降低。SHL 的活性成分可与 γ-分泌酶相互作用。SHL 通过 NOTCH1-MYC 通路诱导 T-ALL 细胞凋亡，可能是治疗 T-ALL 的潜在药物。
