Zhang Simeng, Hua Zhongyan, Ba Gen, Xu Ning, Miao Jianing, Zhao Guifeng, Gong Wei, Liu Zhihui, Thiele Carol J, Li Zhijie
Department of Pediatrics, Shengjing Hospital of China Medical University, Shenyang, China.
Medical Research Center, Liaoning Key Laboratory of Research and Application of Animal Models for Environment and Metabolic Diseases, Shengjing Hospital of China Medical University, #36 Sanhao Street, Heping District, Shenyang, 110004, China.
Cancer Cell Int. 2021 Nov 24;21(1):619. doi: 10.1186/s12935-021-02330-y.
Neuroblastoma (NB) is a common solid malignancy in children that is associated with a poor prognosis. Although the novel small molecular compound Dimethylaminomicheliolide (DMAMCL) has been shown to induce cell death in some tumors, little is known about its role in NB.
We examined the effect of DMAMCL on four NB cell lines (NPG, AS, KCNR, BE2). Cellular confluence, survival, apoptosis, and glycolysis were detected using Incucyte ZOOM, CCK-8 assays, Annexin V-PE/7-AAD flow cytometry, and Seahorse XFe96, respectively. Synergistic effects between agents were evaluated using CompuSyn and the effect of DMAMCL in vivo was evaluated using a xenograft mouse model. Phosphofructokinase-1, liver type (PFKL) expression was up- and down-regulated using overexpression plasmids or siRNA.
When administered as a single agent, DMAMCL decreased cell proliferation in a time- and dose-dependent manner, increased the percentage of cells in SubG1 phase, and induced apoptosis in vitro, as well as inhibiting tumor growth and prolonging survival in tumor-bearing mice (NGP, BE2) in vivo. In addition, DMAMCL exerted synergistic effects when combined with etoposide or cisplatin in vitro and displayed increased antitumor effects when combined with etoposide in vivo compared to either agent alone. Mechanistically, DMAMCL suppressed aerobic glycolysis by decreasing glucose consumption, lactate excretion, and ATP production, as well as reducing the expression of PFKL, a key glycolysis enzyme, in vitro and in vivo. Furthermore, PFKL overexpression attenuated DMAMCL-induced cell death, whereas PFKL silencing promoted NB cell death.
The results of this study suggest that DMAMCL exerts antitumor effects on NB both in vitro and in vivo by suppressing aerobic glycolysis and that PFKL could be a potential target of DMAMCL in NB.
神经母细胞瘤（NB）是儿童常见的实体恶性肿瘤，预后较差。尽管新型小分子化合物二甲基氨基米氏内酯（DMAMCL）已被证明能在某些肿瘤中诱导细胞死亡，但其在NB中的作用尚不清楚。
我们检测了DMAMCL对四种NB细胞系（NPG、AS、KCNR、BE2）的影响。分别使用Incucyte ZOOM、CCK-8检测、Annexin V-PE/7-AAD流式细胞术和Seahorse XFe96检测细胞汇合度、存活率、凋亡率和糖酵解。使用CompuSyn评估药物之间的协同作用，并使用异种移植小鼠模型评估DMAMCL在体内的作用。使用过表达质粒或小干扰RNA上调和下调肝型磷酸果糖激酶-1（PFKL）的表达。
作为单一药物给药时，DMAMCL以时间和剂量依赖性方式降低细胞增殖，增加亚G1期细胞百分比，并在体外诱导凋亡，同时在体内抑制荷瘤小鼠（NGP、BE2）的肿瘤生长并延长生存期。此外，DMAMCL与依托泊苷或顺铂联合使用时在体外发挥协同作用，与单独使用任一药物相比，与依托泊苷联合使用时在体内显示出增强的抗肿瘤作用。机制上，DMAMCL通过降低葡萄糖消耗、乳酸排泄和ATP产生，以及在体外和体内降低关键糖酵解酶PFKL的表达来抑制有氧糖酵解。此外，PFKL过表达减弱了DMAMCL诱导的细胞死亡，而PFKL沉默促进了NB细胞死亡。
本研究结果表明，DMAMCL通过抑制有氧糖酵解在体外和体内对NB发挥抗肿瘤作用，并且PFKL可能是DMAMCL在NB中的潜在靶点。
