二甲双胍对替莫唑胺耐药的胶质母细胞瘤细胞体内、体外抑制作用  

In vivo and in vitro inhibitory effects of metformin on the resistance of glioblastoma cell lines to temozolomide

作　　者：贾迪 林少辉 袁颖 李晨龙[4] 李莎莎 师岩[1] 祝欣萍 彭瑶 李淑艳[1] JIA Di;LIN Shaohui;YUAN Ying;LI Chenlong;LI Shasha;SHI Yan;ZHU Xinping;PENG Yao;LI Shuyan(Department of Medical Technology,Qiqihar Medical University,Heilongjiang Qiqihar 161006,China;Department of Clinical Laboratory,Quanzhou Hospital of Traditional Chinese,Fujian Quanzhou 362005,China;Department of Pathology,Heilongjiang Provincial Hospital,Heilongjiang Harbin 150036,China;Department of Neurosurgery,Harbin Medical University Cancer Hospital,Heilongjiang Harbin 150081,China;Department of Plastic Surgery,the First Affiliated Hospital of Shenzhen University,Guangdong Shenzhen 518025,China;School of Basic Medicine,Heilongjiang University of Chinese Medicine,Heilongjiang Harbin 150040,China)

机构地区：[1]齐齐哈尔医学院医学技术学院,黑龙江齐齐哈尔161006 [2]泉州市中医院检验科,福建泉州362005 [3]黑龙江省医院病理科,黑龙江哈尔滨150036 [4]哈尔滨医科大学附属肿瘤医院神经外科,黑龙江哈尔滨150081 [5]深圳大学附属第一医院整形外科,广东深圳518025 [6]黑龙江中医药大学基础医学院,黑龙江哈尔滨150040

出　　处：《现代肿瘤医学》2024年第21期4054-4061,共8页Journal of Modern Oncology

基　　金：黑龙江省省属高等学校基本科研业务费科研项目(编号:2019-KYYWF-1255)。

摘　　要：目的:探讨二甲双胍对替莫唑胺(temozolomide,TMZ)耐药的胶质母细胞瘤(glioblastoma,GBM)的抑制作用及分子机制,为临床应用二甲双胍治疗GBM耐药提供理论依据。方法:CCK-8法检测二甲双胍对耐药细胞化疗敏感性的影响;流式细胞术检测二甲双胍对耐药细胞凋亡率的影响;细胞克隆形成实验、裸鼠成瘤实验、划痕实验、Transwell侵袭实验、神经球形成实验检测二甲双胍对GBM耐药细胞的克隆形成能力、体内成瘤能力、迁移能力、侵袭能力和干细胞特性的影响;采用甘油三脂含量检测试剂盒检测亲代细胞及耐药细胞在二甲双胍给药前后的甘油三脂含量变化;RT-PCR和Western Blot检测亲代细胞及耐药细胞在二甲双胍给药前后的SREBP1基因和蛋白表达的作用,以及二甲双胍对敲减SREBP1的细胞系耐药性的影响。结果:二甲双胍处理可显著增强LN229-R、U251-R对TMZ的敏感性,并提高其对TMZ的凋亡率。二甲双胍可抑制耐药细胞的克隆形成能力、体内成瘤能力、迁移能力、侵袭能力、干细胞特性和甘油三酯含量(P均<0.05)。二甲双胍降低耐药细胞内SREBP1 mRNA和蛋白表达水平(P均<0.05)。敲减SREBP1可部分逆转耐药性(P<0.05),二甲双胍对敲低SREBP1的耐药细胞无作用(P>0.05)。结论:二甲双胍治疗降低GBM耐药细胞系的克隆形成、增殖、迁移、侵袭和肿瘤干细胞特性,提高耐药细胞对TMZ的凋亡率。二甲双胍可能通过抑制SREBP1降低GBM耐药细胞的甘油三酯含量,从而抑制GBM对TMZ的耐药性。Objective:To explore the inhibitory effect and molecular mechanism of metformin on temozolomide(TMZ)resistance in glioblastoma(GBM),and provide theoretical foundation for clinical application of metformin in the treatment of GBM resistance.Methods:CCK-8 assay was used to detect the effect of metformin on the chemosensitivity of GBM drug-resistant cells.Flow cytometry was used to detect the effect of metformin on the apoptosis rate of GBM drug-resistant cells.Colony formation assay,nude mouse tumorigenesis assay,scratch assay,Transwell invasion assay and neurosphere formation assay were used to detect the effect of metformin on the colony formation ability,tumorigenic ability,migration ability,invasion ability and stem cell characteristics of GBM drug-resistant cells.Triglyceride content detection kits were used to detect the changes of triglyceride in parental cells and GBM drug-resistant cells before and after metformin administration.RT-PCR and Western Blot were used to detect the role of SREBP1 gene and protein expression in the parental cells and GBM drug-resistant cells before and after metformin administration.Results:Metformin treatment could significantly enhance the sensitivity of LN229-R and U251-R to TMZ,and increase the apoptosis rate of drug-resistant cells to TMZ.Metformin could reduce the colony formation ability,the tumor-forming ability in vivo,the migration and invasion ability,the characteristics of tumor stem cells,and the triglyceride content of GBM resistant cells(P all<0.05).Metformin could reduce the expression of SREBP1 mRNA and protein level(P all<0.05)in resistant cells.SREBP1 suppression reversed TMZ resistance of resistant cells(P<0.05),while metformin did not cause significant inhibition in SREBP1-knockdown resistant cells(P>0.05).Conclusion:Metformin treatment reduced the colony formation,proliferation,migration,invasion and tumor stem cell properties of GBM resistant cell lines,and increased the apoptosis rate of GBM resistant cells to TMZ.Metformin may reduce triglyceride content

关 键 词：胶质母细胞瘤 二甲双胍 替莫唑胺 耐药性 

分 类 号：R739.41[医药卫生—肿瘤]