Wang Chong, Li Mengya, Li Lingling, Shen Xiaohui, Liu Yanfang, Wang Shujuan
Department of Hematology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
J Cell Mol Med. 2021 Jan;25(1):272-283. doi: 10.1111/jcmm.15995. Epub 2020 Dec 1.
In this study, we investigated the ability of the Polysaccharide from the Eggs of Strongylocentrotus nudus (SEP) to regulate cellular autophagy and apoptosis in leukaemia cells. Human acute myeloid leukaemia (AML) cells (HL60) and murine AML cells (L1210) treated with SEP were used to assess viability using Cell Counting Kit-8, cytotoxicity by measuring lactate dehydrogenase release, the generation of reactive oxygen species (ROS) by DCFH-DA staining. In addition, we utilized a mouse model of leukaemia in which L1210 cells were injected into DBA/2 mice by sub-axillary injection. Treatment with SEP decreased cell viability, increased in cytotoxicity and increased the release of ROS in a dose-dependent manner. SEP treatment was also associated with the activation of pro-apoptotic proteins cleaved caspase-3, cleaved caspase-9 and cleaved poly (ADP-ribose) polymerase (PARP). Activation of the apoptotic pathway led to the release of cytochrome C (CytoC) into the cytosol of the cell resulting in decreased membrane potential. The effect of SEP treatment was depended on the activation of the nuclear factor kappa-B (NF-κB) signalling pathway as SEP treatment led to an increase in NF-κB phosphorylation, and inhibition of NF-κB signalling using PDTC blocked SEP-mediated activation of apoptosis. Treatment with SEP also prolonged survival time in our leukaemia mouse model and was associated with diminished tumour volume, increased leucocyte and lymphocyte proliferation, promoted pro-inflammatory factor release in serum and enhanced immune function. Taken together, these data suggest that SEP inhibits the progression of leukaemia by initiating mitochondrial dysfunction, autophagy, and apoptosis via the NF-κB signalling pathway.
在本研究中，我们研究了海胆卵多糖（SEP）调节白血病细胞自噬和凋亡的能力。用SEP处理的人急性髓性白血病（AML）细胞（HL60）和小鼠AML细胞（L1210），使用细胞计数试剂盒-8评估细胞活力，通过测量乳酸脱氢酶释放评估细胞毒性，用DCFH-DA染色检测活性氧（ROS）的生成。此外，我们利用了一种白血病小鼠模型，通过腋下注射将L1210细胞注射到DBA/2小鼠体内。SEP处理以剂量依赖的方式降低细胞活力、增加细胞毒性并增加ROS的释放。SEP处理还与促凋亡蛋白裂解的半胱天冬酶-3、裂解的半胱天冬酶-9和裂解的聚（ADP-核糖）聚合酶（PARP）的激活有关。凋亡途径的激活导致细胞色素C（CytoC）释放到细胞质中，导致膜电位降低。SEP处理的效果取决于核因子κB（NF-κB）信号通路的激活，因为SEP处理导致NF-κB磷酸化增加，使用PDTC抑制NF-κB信号通路可阻断SEP介导的凋亡激活。SEP处理还延长了我们白血病小鼠模型的生存时间，并与肿瘤体积减小、白细胞和淋巴细胞增殖增加、血清中促炎因子释放促进以及免疫功能增强有关。综上所述，这些数据表明SEP通过NF-κB信号通路引发线粒体功能障碍、自噬和凋亡来抑制白血病的进展。
