皖南尖吻蝮蛇蛇毒抑瘤组分Ⅰ促进Erastin诱导人口腔鳞癌Hsc3细胞发生铁死亡的相关机制  

Promotion mechanism of AAVC-I for Erastin induced ferroptosis in human oral squamous cell carcinoma cell line Hsc3

作　　者：陶桃 陶志豪 邢新 张方 王之恒 柴琳 Tao Tao;Tao Zhihao;Xing Xin;Zhang Fang;Wang Zhiheng;Chai Lin(School of Stomatology,Wannan Medical College,Wuhu,Anhui 241000,China;Department of Oral and Maxillofacial Surgery,the First Affiliated Hospital of Wannan Medical College,Wuhu,Anhui 241000,China)

机构地区：[1]皖南医学院口腔医学院,安徽芜湖241000 [2]皖南医学院第一附属医院口腔颌面外科

出　　处：《齐齐哈尔医学院学报》2024年第9期801-807,共7页Journal of Qiqihar Medical University

基　　金：安徽省高校科学研究重大项目(KJ2021ZD0103);芜湖市科技局应用基础研究项目(2022jc68);皖南医学院中青年科研基金(WK202202;WK2023ZQNZ41);活性生物大分子安徽省重点实验室开放课题(LAB202206)。

摘　　要：目的 研究皖南尖吻蝮蛇蛇毒抑瘤组分Ⅰ(anti-tumor component Ⅰ from Agkistrodon acutus venom, AAVC-Ⅰ)是否能够促进Erastin诱导口腔鳞癌细胞铁死亡的水平并初步探索相关机制。方法 通过CCK8检测细胞活力,通过流式细胞仪检测细胞中活性氧、脂质氧化以及线粒体膜电位水平,通过Western-blot检测细胞内胱氨酸谷氨酸转运受体(system XC-,xCT)、谷胱甘肽过氧化物酶4(Glutathione Peroxidase 4,GPX4)蛋白表达水平,通过还原性谷胱甘肽(Glutathione, GSH)、丙二醛(Malondialdehyde, MDA)试剂盒检测细胞内GSH、MDA水平。结果 经Erastin和浓度为2μg/ml的AAVC-Ⅰ联合处理后,细胞活力相比于单独的Erastin处理出现了明显下降(P<0.05),并且可以被铁死亡抑制剂Fer-1所逆转(P>0.05),同时联合处理组细胞中活性氧、脂质氧化水平出现了明显上升(P<0.05),而线粒体膜电位、GSH水平明显下降(P<0.05),此外xCT、GPX4蛋白表达被抑制(P<0.05)。结论 浓度为2μg/ml的AAVC-Ⅰ可以促进Erastin诱导口腔鳞癌细胞铁死亡的水平,其机制可能是通过抑制xCT/GPX4使细胞抗氧化能力下降,氧化应激水平增加。Objective To examine if anti-tumor component I from Agkistrodon acutus venom(AAVC-I)can enhance Erastin induced ferroptosis in oral squamous cell carcinoma cells and to preliminarily investigate the underlying mechanisms.Methods Cell viability was assessed by using the CCK8 assay,while reactive oxygen species,lipid oxidation,and mitochondrial membrane potential were measured via flow cytometry.The expression levels of system XC-(xCT)and Glutathione Peroxidase 4(GPX4)proteins were determined by Western blot analysis,and the concentrations of Glutathione(GSH)and Malondialdehyde(MDA)in cells were quantified by using assay kits.Results Treatment with Erastin and AAVC-I at a concentration of 2μg/mL significantly reduced cell viability compared to Erastin alone(P<0.05),an effect that was reversible by the ferroptosis inhibitor Fer-1(P>0.05).Additionally,the combined treatment resulted in a marked increase in reactive oxygen species and lipid oxidation levels(P<0.05),alongside a significant decrease in mitochondrial membrane potential and GSH levels(P<0.05).The expression of xCT and GPX4 proteins was also inhibited(P<0.05).Conclusions AAVC-I at a concentration of 2μg/ml can enhance Erastin induced ferroptosis in oral squamous cell carcinoma cells.Its mechanism may involve diminishing the cell's antioxidant capacity and elevating oxidative stress levels by inhibiting the xCT/GPX4 pathway.

关 键 词：铁死亡 蛇毒 口腔鳞癌 氧化应激 

分 类 号：R246.83[医药卫生—针灸推拿学]