膜联蛋白ANXA5调节黑色素瘤细胞铁死亡及其机制研究  

Mechanisms of annexin A5 regulating ferroptosis sensitivity in melanoma cells

作　　者：王嘉慧 李璐琪 张悦 张肖文 张晶[1] WANG Jiahui;LI Luqi;ZHANG Yue;ZHANG Xiaowen;ZHANG Jing(School of Life Sciences,Nanjing University,Nanjing 210023,China)

机构地区：[1]南京大学生命科学学院,江苏南京210023

出　　处：《南京医科大学学报(自然科学版)》2025年第2期157-164,共8页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(32250016)。

摘　　要：目的:探究黑色素瘤中膜联蛋白A5(annexinA5,ANXA5)表达与铁死亡的相关性。方法:利用The Human Protein Atlas数据分析ANXA5在黑色素瘤中的表达情况,并采用人黑素瘤组织芯片进行免疫组化验证,再以黑色素瘤细胞B16F10为研究对象,通过CRISPR-Cas9技术构建ANXA5敲除株,通过CCK-8及乳酸脱氢酶(lactate dehydrogenase,LDH)检测敲除ANXA5对B16F10铁死亡效应的影响,进一步通过RNA-seq寻找ANXA5调节铁死亡可能的靶分子并验证,最后在不同的黑色素瘤细胞中检测内源性ANXA5表达水平,并确认ANXA5对铁死亡敏感性的调节作用。结果:ANXA5在黑色素瘤组织中异常高表达,敲除或敲低ANXA5能促进黑色素瘤细胞对铁死亡的敏感性。酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4,ACSL4)作为铁死亡关键调节分子,与ANXA5表达呈负相关性,过表达ANXA5能降低ACSL4水平,表现出对铁死亡的抵抗性。结论:在黑色素瘤细胞中,敲低ANXA5能够上调ACSL4水平,增强黑色瘤细胞对铁死亡的敏感性,提示ANXA5有望成为黑色素瘤铁死亡治疗的潜在靶点。Objective:To explore the correlation between the expression of annexin A5(ANXA5)and ferroptosis in melanoma.Methods:The expression of ANXA5 in melanoma was analyzed using The Human Protein Atlas data and further validated by immunohistochemistry on a human melanoma tissue microarray.The B16F10 melanoma cell line was used as the research model,where ANXA5 knockout cell lines were generated using CRISPR⁃Cas9 technology.The effect of ANXA5 knockout on ferroptosis in B16F10 cells was assessed using CCK⁃8 and lactate dehydrogenase(LDH)assays.RNA⁃seq was further performed to identify the possible target molecules of ANXA5 related to ferroptosis,followed by validation.Additionally,the endogenous expression of ANXA5 in different melanoma cell lines was measured to confirm its regulatory role in ferroptosis sensitivity.Results:ANXA5 was aberrantly overexpressed in melanoma tissues,and knockingdown or knockingout ANXA5 promoted the sensitivity to ferroptosis in melanoma cells.Acyl⁃CoA synthetase long⁃chain family member 4(ACSL4)as a key regulator of ferroptosis,was negatively correlated with ANXA5 expression.Overexpression of ANXA5 reduced the level of ACSL4 and showed resistance to ferroptosis.Conclusion:In melanoma cells,downregulation of ANXA5 enhanced the sensitivity to ferroptosis via upregulating ACSL4,suggesting that ANXA5 may be a potential target of ferroptosis⁃based therapy in melanoma.

关 键 词：黑色素瘤 铁死亡 膜联蛋白A5 ASCL4 

分 类 号：R739.5[医药卫生—肿瘤]