KIAA1199调控铁死亡促进骨肉瘤细胞增殖的机制探究  

Mechanism of KIAA1199 promoting osteosarcoma cell proliferation by regulating ferroptosis

作　　者：龚辉松 程军 邓幼文 Gong Huisong;Cheng Jun;Deng Youwen(Department of Spine Surgery,Guangdong Hospital of Integrated Traditional Chinese and Western Medicine,Foshan 528200,China)

机构地区：[1]广东省中西医结合医院脊柱骨科,广东佛山528200 [2]中南大学湘雅三医院脊柱外科

出　　处：《海军医学杂志》2025年第4期392-400,共9页Journal of Navy Medicine

基　　金：佛山市自筹经费类科技创新项目(2220001003886)。

摘　　要：目的探讨细胞迁移诱导蛋白(KIAA1199)对骨肉瘤细胞增殖的影响及机制。方法在KIAA1199高表达的骨肉瘤细胞系(HOS和U2OS)中将KIAA1199基因敲除或构建KIAA1199过表达细胞,运用RT⁃PCR和Western blot验证KIAA1199表达水平,运用CCK⁃8实验、细胞克隆形成实验检测KIAA1199对细胞增殖的影响,CCK⁃8、细胞活力实验检测KIAA1199促进骨肉瘤细胞增殖的相关机制,BODIPY 581/591探针检测KIAA1199对细胞脂质过氧化的影响,RT⁃PCR分析KIAA1199对细胞前列腺素内过氧化物合酶2(PTGS2)的影响,Western blot检测KIAA1199敲除对铁死亡相关蛋白谷胱甘肽过氧化物酶4(GPX4)、溶质载体家庭7成员11(SLC7A11)、二氢乳清酸脱氢酶(DHODH)、铁死亡抑制蛋白1(FSP1)的影响。结果KIAA1199敲除会抑制骨肉瘤细胞增殖,这种抑制效应主要通过促进骨肉瘤细胞铁死亡发挥功能;铁死亡抑制剂Fer⁃1可逆转KIAA1199敲除造成的增殖抑制现象;KIAA1199敲除的骨肉瘤细胞对铁死亡诱导剂Erastin更敏感,脂质过氧化水平和PTGS2 mRNA表达水平明显增高;KIAA1199敲除会造成GPX4和SLC7A11蛋白表达水平降低、还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)比值降低。将KIAA1199过表达可促进骨肉瘤细胞增殖,使细胞对Erastin更耐受,脂质过氧化水平和PTGS2 mRNA表达水平降低,GPX4和SLC7A11蛋白表达水平增加。结论KIAA1199通过SLC7A11/GSH/GPX4信号轴抑制铁死亡,进而促进骨肉瘤细胞增殖。Objective To investigate the effect and mechanism of cell migration⁃induced protein(KIAA1199)on the proliferation of osteosarcoma cells.Methods The KIAA1199 gene was knocked down or overexpressed in osteosarcoma cell lines(HOS and U2OS)with high KIAA1199 expression.The expression of KIAA1199 was confirmed by RT⁃PCR and Western blotting.CCK⁃8 and colony formation assays were used to determine the effect of KIAA1199 on cell proliferation.CCK⁃8 and cell viability assays were used to confirm that KIAA1199 promoted osteosarcoma cell proliferation by inhibiting ferroptosis.The BODIPY 581/591 probe was used to investigate the effect of KIAA1199 on cell lipid peroxidation.The effect of KIAA1199 on PTGS2 was investigated by RT⁃PCR,and the level of ferroptosis related proteins(GPX4,SLC7A11,DHODH,and FSP1)were determined by Western blotting.Results KIAA1199 knockdown inhibited osteosarcoma cell proliferation,which mainly worked by promoting ferroptosis in osteosarcoma cells.Ferrostatin⁃1(Fer⁃1,a ferroptosis inhibitor)could reverse the proliferation inhibition caused by KIAA1199 knockdown.Osteosarcoma cells with KIAA1199 knockdown were more sensitive to ferroptosis inducer Erastin,and their lipid peroxidation and PTGS2 mRNA increased significantly.KIAA1199 knockdown resulted in lower expression of GPX4 and SLC7A11 proteins and a lower GSH/GSSG ratio.On the other hand,KIAA1199 overexpression promoted osteosarcoma cell proliferation,made cells more tolerant to Erastin,significantly reduced lipid peroxidation and PTGS2 mRNA levels,and obviously increased the protein expression levels of GPX4 and SLC7A11.Conclusion KIAA1199 inhibits ferroptosis through SLC7A11/GSH/GPX4 pathway,thereby promoting osteosarcoma cell proliferation.

关 键 词：细胞迁移诱导蛋白 骨肉瘤 细胞增殖 铁死亡 

分 类 号：R738.1[医药卫生—肿瘤]