消皮素D基因高表达同胶质瘤不良预后相关  

High expression of Gasdermin D gene is associated with poor prognosis of gliomas

作　　者：蒲亚陆 周文科[1,2] 梁文 李富光 王涛[1] 时志超 杨如[1,2] Pu Yalu;Zhou Wenke;Liang Wen;Li Fuguang;Wang Tao;Shi Zhichao;Yang Ru(Department of Neurosurgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China;Life Science Research Center of the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China)

机构地区：[1]新乡医学院第一附属医院神经外科,卫辉453100 [2]新乡医学院第一附属医院生命科学研究中心,卫辉453100

出　　处：《中华实验外科杂志》2022年第6期1161-1164,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探究消皮素D(GSDMD)在胶质瘤发生发展过程中的意义和体外实验中对胶质瘤细胞系增殖、迁移的影响及其分子机制。方法利用生物信息学的方法分析胶质瘤中GSDMD基因的表达特点, 及GSDMD表达水平高低同临床预后的关系。采用实时荧光定量聚合酶链式反应(qRT-PCR)法检测人胶质瘤细胞U251MG、U87MG、LN229及正常胶质细胞HMC3中GSDMD的表达;运用慢病毒GSDMD基因敲除载体(LV-shGSDMD)与慢病毒空载对照(LV-NC)转染胶质瘤细胞LN229, 并筛选稳转株LN229-NC和LN229-shGSDMD;应用qRT-PCR检测GSDMD mRNA的表达水平;细胞计数试剂盒(CCK-8)法检测LN229-NC、LN229-shGSDMD细胞的增殖能力;采用Transwell检测LN229-NC、LN229-shGSDMD细胞的迁移能力;采用蛋白质印迹法(Western blot)检测LN229-NC、LN229-shGSDMD细胞GSDMD、Nod样受体家族包含pyrin结构域蛋白3(NLRP3)、半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-1、白细胞介素(IL)-1β蛋白的表达情况, 两组间的比较采用双尾Studentt检验。结果在癌症基因组图谱(TCGA)数据库及基因组织表达(GTEx)数据库中, 胶质瘤组GSDMD基因表达水平高于对照组中的正常组织(3.62±0.04比2.83±0.01, t=24.44, P<0.05), 胶质瘤中GSDMD高表达组中位生存时间短于低表达组[2.1年比7.9年, 风险比(HR)=4.5, P<0.05];胶质瘤中GSDMD、IL-1β、Caspase-1表达水平高于正常组织, 且随胶质瘤等级升高而上升。胶质瘤中NLRP3的表达水平同样显著高于正常组织, 但同胶质瘤的等级没有明显关联。在细胞实验中, GSDMDmRNA在胶质瘤细胞系U251MG、U87MG、LN229中的表达量显著高于正常胶质细胞HMC3(4.04±0.35、4.76±0.21、6.29±0.28比1.00±0.07, P<0.05), 而LN229高于其他胶质瘤细胞系(F=200.2, P<0.05);基因敲除后, LN229-shGSDMD组较LN229-NC组细胞中GSDMD mRNA的表达量下降(0.15±0.04比1.00±0.01, t=14.32, P< 0.05);CCK-8实验中, LN229-shGSDMD组细胞吸光度(A)值曲线在LN229-NC组细胞下方, Objective To explore the significance of Gasdermin-D(GSDMD)in the occurrence and development of GBM and its effect on the proliferation and migration of glioma cell line in vitro and its possible molecular mechanism.Methods Bioinformatics method was used to analyze the expression characteristics of GSDMD gene in gliomas and the relationship between GSDMD expression level and clinical prognosis.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of GSDMD in human glioma cells U251MG,U87MG,LN229(human glioma cells)and normal glial cells HMC3(human glial cells).Lentivirus GSDMD gene knockout vector LV-shGSDMD and lentivirus empty LV-NC were used to transfect glioma cell line LN229,and stable transformants LN229-NC and LN229-shGSDMD were selected to detect the expression of GSDMDmRNA by qRT-PCR.The proliferation of LN229-NC and LN229-shGSDMD cells was detected by cell counting kit-8(CCK-8)assay,and the migration ability of LN229-NC and LN229-shGSDMD cells was detected by Transwell.The expression of GSDMD,NLR family,pyrin domain containing 3(NLRP3),cysteinyl aspartate-specific protease(Caspase)-1 and interleukin(IL)-1βin LN229-NC and LN229-shGSDMD cells was detected by Western blotting.Results In cancer genome map(the cancer genome atlas,TCGA)database and gene tissue expression(genotype-tissue expression,GTEx)database,the expression level of GSDMD gene in glioma group was higher than that in control group(3.62±0.04 vs.2.83±0.01,t=24.44,P<0.05).The median survival time in high GSDMD expression group in glioma was shorter than that in low expression group(2.1 years vs.7.9 years,HR=4.5,P<0.05).The expression levels of GSDMD,IL-1βand Caspase-1 in gliomas were higher than those in normal tissues,and increased with the increase of glioma grade.The expression level of NLRP3 in gliomas was also significantly higher than that in normal tissues,but it was not significantly related to the grade of gliomas.In cell experiment,the expression of GSDMD mRNA in glioma cell lines

关 键 词：胶质瘤 增殖 迁移 焦亡 

分 类 号：R739.41[医药卫生—肿瘤]