干扰circRNA_002178抑制胶质瘤细胞恶性生物学行为  

Interfering with circRNA_002178 inhibits malignant biological behavior of glioma cells

作　　者：高雪亮[1] 张皓翔 程建业[1] 贾培雷[1] 赵亚鹏[1] GAO Xueliang;ZHANG Haoxiang;CHENG Jianye;JIA Peilei;ZHAO Yapeng(Department of Neurosurgery,Hebei Hospital of Traditional Chinese Medicine,Shijiazhuang 050000,China)

机构地区：[1]河北省中医院神经外科,石家庄050000 [2]广州中医药大学中西医结合专业,广州510000

出　　处：《中国免疫学杂志》2022年第11期1338-1342,1348,共6页Chinese Journal of Immunology

基　　金：河北省中医药管理局科研计划项目(2019028)。

摘　　要：目的:探讨干扰circRNA_002178对胶质瘤细胞增殖、凋亡、侵袭、炎症反应的影响。方法:RT-qPCR检测临床样本癌组织和癌旁组织中circRNA_002178的表达;转染分组后采用RT-qPCR检测不同转染序列中circRNA_002178的表达,筛选干扰效果最佳序列组;CCK8法检测细胞增殖率;克隆形成法检测细胞形成率;流式检测细胞凋亡水平;Western blot检测凋亡相关蛋白的表达水平;RT-qPCR检测VEGF mRNA表达水平;Western blot检测VEGF蛋白表达水平;Transwell检测侵袭;ELISA检测血清炎症因子含量。结果:胶质瘤组织circRNA_002178表达水平明显高于癌旁组织,选用胶质瘤细胞U-251进行后续实验,选择干扰效果最为显著的circ_002178-shRNA2组为后续实验干扰组。与shRNA-NC组相比,circ_002178-shRNA2组胶质瘤细胞凋亡率与克隆形成率明显降低;胶质瘤细胞凋亡率及Bax/Bcl-2蛋白表达明显升高;胶质瘤细胞VEGF及其VEGF mRNA表达明显降低,侵袭细胞数量明显减少;促炎因子IL-6、iNOS水平降低,抗炎因子IL-10水平升高。结论:干扰circRNA_002178可抑制胶质瘤细胞的增殖、侵袭、炎症反应,诱导其凋亡,并有效抑制胶质瘤细胞恶性生物学行为,有望成为治疗胶质瘤的新靶点。Objective:To investigate the effect of interference circRNA_002178 on proliferation,apoptosis,invasion and inflammation of glioma cells.Methods:RT-qPCR was used to detect expressions of circRNA_002178 in cancer tissues and adjacent tissues of clinical samples;RT-qPCR was used to detect expressions of circRNA_002178 in different transfected sequences after transfection grouping,and the sequence group with the best interference effect was screened;CCK8 method was used to detect cell proliferation rate;clone formation method was used to detect cell formation rate;flow cytometry was used to detect apoptosis level;Western blot was used to detect apoptosis-related protein expression;RT-qPCR was used to detect VEGF mRNA expression;Western blot was used to detect VEGF protein expression;Transwell was used to detect invasion;ELISA was used to detect serum inflammatory factor content.Results:Expression level of circRNA_002178 in glioma tissue was significantly higher than that in adjacent tissues.Glioma cell U-251 was used for follow-up experiments.circ_002178-shRNA2 group with the most significant interference effect was selected as interference group for subsequent experiments.Compared with shRNA-NC group,circ_002178-shRNA2 group glioma cell apoptosis rate and clone formation rate were significantly reduced;glioma cell apoptosis rate and Bax/Bcl-2 protein expression were significantly increased;expressions of VEGF and VEGF mRNA were significantly reduced in glioma cells,and the number of invaded cells was significantly reduced;levels of pro-inflammatory factors IL-6 and iNOS were reduced,while level of anti-inflammatory factor IL-10 was increased.Conclusion:Interfering with circRNA_002178 can inhibit proliferation,invasion and inflammation of glioma cells,and induces their apoptosis,and can effectively inhibit malignant biological behavior of glioma cells,which is expected to become a new target for the treatment of glioma.

关 键 词：胶质瘤 circRNA_002178 侵袭 TRANSWELL 血管内皮生长因子 

分 类 号：R739.42[医药卫生—肿瘤]