miR-330-3p/SLC7A11调节口腔鳞状细胞癌细胞的增殖、凋亡和铁死亡的功能及机制  

The Function and Mechanism of MiR-330-3p/SLC7A11 in Regulating Proliferation,Apoptosis and Ferroptosis in Oral Squamous Carcinoma cells

作　　者：靳昊[1] 张华[1] 吕越[1] 步恒彬 JIN Hao;ZHANG Hua;LV Yue;BU Hengbin(Department of Stomatology,Baoding Second Central Hospital,Baoding 072750;Department of Stomatology,Zhuozhou Hospital of Traditional Chinese Medicine,Zhuozhou 072750)

机构地区：[1]保定市第二中心医院口腔科,保定072750 [2]涿州市中医院口腔科,涿州072750

出　　处：《湖南师范大学学报(医学版)》2024年第6期29-34,145,共7页Journal of Hunan Normal University(Medical Sciences)

基　　金：2021年保定市科技计划“SLC7A11-AS1调控miR-429对口腔鳞癌细胞CAL-27细胞增殖、凋亡及迁移的影响”(2141ZF019);2020年河北省卫健委重点科技研究计划“miR-330-3pSLC7A11表达在口腔鳞状细胞癌铁死亡中的作用”(20201256)

摘　　要：目的:探讨miR-330-3p/SLC7A11在干扰素-γ(interferon-γ,IFN-γ)调控口腔鳞状细胞癌铁死亡中的功能和机制。方法:培养人口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞系,分为对照组和IFN-γ组。CCK8、集落形成测定、细胞侵袭实验和流式细胞术检测细胞活力、迁徙、侵袭和凋亡。qPCR和蛋白质印迹法测量OSCC细胞中miR-330-3p、SLC7A11表达,靶向位点由TargetScan预测,并通过双荧光素酶报告测定法确认,通过操纵miR-330-3p和SLC7A11在OSCC细胞系中的表达水平。铁、ROS水平分析和裸鼠研究检测铁死亡和IFN-γ调控OSCC细胞在体内的生长。结果:与对照组比较,IFN-γ显着降低了OSCC细胞活力,抑制了OSCC细胞的增殖、迁移和侵袭;IFN-γ抑制了OSCC细胞中铁死亡阴性调节因子的水平,包括GPX4和SLC7A11,IFN-γ诱导的ROS、铁和Fe^(2+)在OSCC细胞中积累。通过荧光素酶报告法验证靶向关系,miR-330-3p可有效抑制OSCC细胞中SLC7A11水平。miR-330-3p模拟物增加了OSCC细胞迁移,并降低了细胞凋亡和铁死亡。而裸鼠的致瘤性测定表明,IFN-γ抑制了OSCC细胞在体内的生长。结论:miR-300-3p降低SLC7A11的表达促进IFN-γ诱导的OSCC细胞的增殖和侵袭,抑制细胞凋亡和铁死亡。Objective To investigate the function and mechanism of miR-330-3p/SLC7A11 in the regulation of proliferation,apoptosis and ferroptosis in oral squamous cell carcinoma by interferon-γ.Methods Human OSCC cell lines were cultured and divided into control group and IFN-γgroup.Cell viability,migration,invasion,and apoptosis were measured by CCK8,colony formation assay,cell invasion assay,and flow cytometry.The expression of miR-330-3p and SLC7A11 in OSCC cells was measured by qPCR and Western blotting.The target sites of miR-330-3p were predicted by TargetScan and confirmed by dual luciferase reporting assay,which manipulated the expression levels of miR-330-3p and SLC7A11 in OSCC cell lines.Ferroptosis and IFN-γregulation of OSCC cell growth in vivo were detected by iron and ROS level analysis and nude mouse study.Results Compared with the control group,IFN-γsignificantly reduced OSCC cell viability and inhibited the proliferation,migration and invasion of OSCC cells.IFN-γinhibited levels of negative regulators of iron apoptosis in OSCC cells,including GPX4 and SLC7A11,IFN-γ-induced ROS,and accumulation of iron and Fe^(2+)in OSCC cells.The targeting relationship was verified by luciferase reporting method,and miR-330-3p effectively inhibited SLC7A11 levels in OSCC cells.MiR-330-3p mimics increased OSCC cell migration and decreased apoptosis and ferroptosis.Tumogenicity in nude mice showed that IFN-γinhibited OSCC cell growth in vivo.Conclusion MiR-300-3p inhibits the expression of SLC7A11 to promote IFN-γ-induced proliferation and invasion of OSCC cells,and inhibit cell apoptosis and ferroptosis.

关 键 词：miR-300-3p SLC7A11 干扰素-Γ 口腔鳞状细胞癌 铁死亡 

分 类 号：R739.8[医药卫生—肿瘤]