Department of Basic Medicine, Changzhi Medical College, Changzhi, Shanxi Province, China.
Changzhi Maternal and Child Health Care Hospital, Changzhi, Shanxi Province, China.
Comput Math Methods Med. 2022 Jul 8;2022:2296574. doi: 10.1155/2022/2296574. eCollection 2022.
The focus of this research was to look at the effects of the combination of the lentinan (LNT) and oxaliplatin (Oxa) on the apoptosis of human esophageal cancer cells, as well as the underlying mechanism.
LNT and Oxa were used to treat EC-109 human esophageal cancerous cells at various doses, and the cell survival rate was measured using the Cell Counting Kit-8 (CCK-8) assay. In addition, 24 h after treatment of EC-109 cells with a combination of LNT and Oxa, flow cytometry was used to analyze their apoptotic effect on these cells. Additionally, LNT on EC-109 cell apoptotic upshot was assessed via measuring the consequence of LNT on the mRNA and protein expression levels pertaining to immunogenic cell death factors CALR, HSP90, and HSP70 by qPCR (quantitative real-time polymerase chain reaction) and western blot analysis, correspondingly.
Cell proliferation was inhibited only when EC-109 cells were added with LNT at 1,200 g/mL to the maximum concentrations, but the combination of LNT and Oxa at a low dose (800 g/mL and 20 M, respectively) significantly increased their sensitivity to Oxa and reduced their proliferation ( < 0.05), and their apoptosis was significantly increased by LNT ( < 0.05). The immunogenic cell death-related genes CALR, HSP90, and HSP70 had dramatically enhanced mRNA and protein expression levels after therapy with a combination of LNT and Oxa ( < 0.05).
These data imply that LNT increases the susceptibility of esophageal cancerous cells to Oxa by driving EC-109 cells to display immunogenic death. Therefore, LNT combined with Oxa may be an effective method in esophageal cancer management.
本研究旨在观察香菇多糖(LNT)和奥沙利铂(Oxa)联合作用对人食管癌细胞凋亡的影响及其机制。
用不同剂量的 LNT 和 Oxa 处理 EC-109 人食管癌细胞,用细胞计数试剂盒-8(CCK-8)法测定细胞存活率。此外,用 LNT 和 Oxa 联合处理 EC-109 细胞 24 h 后,用流式细胞术分析它们对这些细胞的凋亡作用。此外,通过 qPCR(实时定量聚合酶链反应)和 Western blot 分析相应地评估 LNT 对免疫原性细胞死亡因子 CALR、HSP90 和 HSP70 的 mRNA 和蛋白表达水平的影响,来评估 LNT 对 EC-109 细胞凋亡的影响。
仅当 EC-109 细胞中 LNT 的最大浓度达到 1200μg/ml 时,细胞增殖才受到抑制,但 LNT 和 Oxa 的低剂量(分别为 800μg/ml 和 20μM)联合使用可显著提高 Oxa 的敏感性,降低细胞增殖(<0.05),并显著增加 LNT 的细胞凋亡(<0.05)。联合使用 LNT 和 Oxa 治疗后,免疫原性细胞死亡相关基因 CALR、HSP90 和 HSP70 的 mRNA 和蛋白表达水平显著增强(<0.05)。
这些数据表明,LNT 通过诱导 EC-109 细胞发生免疫原性死亡,增加食管癌细胞对 Oxa 的敏感性。因此,LNT 联合 Oxa 可能是治疗食管癌的有效方法。