诱导免疫原性细胞死亡（ICD）过程可能是卵巢癌（OC）的重要抗肿瘤策略。二甲双胍（Met）已被证明在 OC 中具有抗肿瘤作用，但其是否介导 ICD 抑制 OC 过程尚不清楚。用 Met 处理人 OC 细胞系（SKOV3 和 A2780）。从志愿者的外周血单核细胞中分离出树突状细胞 (DC) 和 CD8 T 细胞。采用细胞计数试剂盒8法测定细胞活力，免疫荧光染色检测细胞膜和细胞内钙网蛋白（CRT）的百分比。通过流式细胞术评估CRT水平、DC成熟和效应细胞活化。使用相应试剂盒检测IL-10和IFN-γ的水平以及HMGB1和ATP的释放量。通过蛋白质印迹分析检测热休克蛋白70/90（HSP70/90）和AMPKα的蛋白水平，通过实时定量PCR检测CD80、CD86、IL-10和IFN-γ的mRNA水平。利用集落形成测定来评估细胞的细胞毒性。构建小鼠移植瘤模型以评估Met对OC肿瘤生长的影响，并采用免疫组织化学染色分析小鼠肿瘤组织中的CD80和CD86细胞。我们的数据表明，Met 抑制 OC 细胞活力并诱导 CRT 暴露。此外，Met还可以促进HMGB1和ATP的释放，并诱导DC成熟。体内实验表明，Met 通过激活抗肿瘤免疫反应来抑制 OC 肿瘤的生长。此外，Met激活AMPK通路，沉默AMPK通路逆转了Met对CRT暴露以及OC细胞中HMGB1和ATP释放的促进作用。总之，Met通过激活AMPK通路在OC中诱导ICD介导的免疫破坏，表明Met可能用于OC的免疫治疗。版权所有©2024。由Elsevier Inc.出版。

Inducing immunogenic cell death (ICD) process may be an important antitumor strategy in ovarian cancer (OC). Metformin (Met) has been shown to have antitumor effects in OC, but whether it mediates the ICD to inhibit OC process is unclear. Human OC cell lines (SKOV3 and A2780) were treated with Met. Dendritic cell (DC) and CD8+T cells were isolated from the peripheral blood mononuclear cells of volunteers. Cell counting kit 8 assay was used to measure cell viability, and immunofluorescence staining was performed to detect the percentages of membrane and intracellular calreticulin (CRT). CRT level, DC maturation and effector cell activation were evaluated by flow cytometry. The levels of IL-10 and IFN-γ, as well as the releasements of HMGB1 and ATP, were detected using corresponding kits. The protein levels of heat shock protein 70/90 (HSP70/90) and AMPKα were tested by western blot analysis, and the mRNA levels of CD80, CD86, IL-10, and IFN-γ were measured by quantitative real-time PCR. Colony formation assay was utilized for assessing cell cytotoxicity. Mice transplanted tumor model was constructed to assess the effect of Met on OC tumor growth, and immunohistochemistry staining was used to analyze CD80+ and CD86+ cells in mice tumor tissues. Our data showed that Met inhibited OC cell viability and induced CRT exposure. Besides, Met could promote the release of HMGB1 and ATP, as well as induce DC maturation. In vivo experiments suggested that Met restrained OC tumor growth via activating antitumor immune response. Moreover, Met activated AMPK pathway, and silenced AMPK pathway reversed the promoting effect of Met on CRT exposure and the releasements of HMGB1 and ATP in OC cells. In conclusion, Met induced ICD-mediated immune destruction in OC via activating AMPK pathway, indicating that Met might be used in the immunotherapy of OC.Copyright © 2024. Published by Elsevier Inc.