Department of Thoracic Surgery, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University, Qingdao, China.
Department of Thoracic Surgery, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
Anat Rec (Hoboken). 2021 Feb;304(2):323-332. doi: 10.1002/ar.24414. Epub 2020 May 24.
Esophageal squamous cell carcinoma (ESCC) presents a common human malignancy in the digestive system. We aimed to explore the critical effects of alpinumisoflavone (AIF) on ESCC in vitro and in vivo. The cell counting kit-8 assay was used to determine cell viability. Colony formation assay was employed to examine the effect of AIF on the long-term growth of ESCC cells. Cell apoptosis was determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. Cell morphologies were observed by light microscopy. The enzyme-linked immunosorbent assay was performed to examine the lactate dehydrogenase release from AIF-treated cells. Immunofluorescent labeling was utilized to examine AIF-induced GSDME expression. Western blot was employed to determine the expression levels of the associated proteins. Immunohistochemistry was performed to determine the localization and expression of the associated proteins in mice tumor tissues. AIF inhibited ESCC cell viability and suppressed cell growth in a dose- and time-dependent fashion. Results showed that AIF promoted apoptosis in ESCC cells. Meanwhile, our results also showed that AIF triggered pyroptotic cell death in ESCC, which was mediated by gasdermin E (GSDME) cleavage. In addition, our experiments provided experimental evidence that AIF-induced GSDME cleavage was dependent on caspase-3 activation. Moreover, the inhibition of GSDSE by knockdown was able to switch the form of cell death from pyroptosis to apoptosis. Furthermore, the results from the xenograft animal model also supported our findings in vitro that AIF was able to promote GSDME-mediated pyroptotic cell death in ESCC. AIF inhibited ESCC growth in vitro and in vivo by triggering GSDME-mediated pyroptotic cell death, which is dependent on caspase-3 activation.
食管鳞状细胞癌 (ESCC) 是消化系统中常见的人类恶性肿瘤。我们旨在探讨白杨素 (AIF) 在 ESCC 体外和体内的关键作用。细胞计数试剂盒-8 测定法用于确定细胞活力。集落形成测定法用于研究 AIF 对 ESCC 细胞长期生长的影响。通过流式细胞术和末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记法测定细胞凋亡。通过光镜观察细胞形态。酶联免疫吸附测定法用于检测 AIF 处理细胞中乳酸脱氢酶的释放。免疫荧光标记用于检测 AIF 诱导的 GSDME 表达。Western blot 用于确定相关蛋白的表达水平。免疫组织化学用于确定相关蛋白在小鼠肿瘤组织中的定位和表达。AIF 抑制 ESCC 细胞活力并呈剂量和时间依赖性抑制细胞生长。结果表明，AIF 促进 ESCC 细胞凋亡。同时，我们的结果还表明，AIF 通过 gasdermin E (GSDME) 切割触发 ESCC 中的细胞焦亡。此外，我们的实验提供了实验证据，表明 AIF 诱导的 GSDME 切割依赖于 caspase-3 激活。此外，通过敲低抑制 GSDSE 能够将细胞死亡形式从细胞焦亡转变为细胞凋亡。此外，异种移植动物模型的结果也支持了我们在体外的发现，即 AIF 能够通过触发 GSDME 介导的细胞焦亡来促进 ESCC 中的细胞焦亡。AIF 通过触发 GSDME 介导的细胞焦亡来抑制 ESCC 的体外和体内生长，这依赖于 caspase-3 的激活。
