PDIA4基因沉默对胶质瘤细胞凋亡的影响  

Effect of protein disulfide isomerase family A member 4 silencing on apoptosis of glioma cell

作　　者：朱传华 谢琳[1] 刘安民[1] ZHU Chuan-hua;XIE Lin;LIU An-min(Department of Neurosurgery,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510120,China)

机构地区：[1]中山大学孙逸仙纪念医院神经外科,广州510120

出　　处：《岭南现代临床外科》2021年第2期177-181,共5页Lingnan Modern Clinics in Surgery

基　　金：广州市科技计划项目(201803010045)。

摘　　要：目的探讨蛋白二硫键异构酶家族成员4(PDIA4)基因在胶质瘤细胞中的表达及对细胞凋亡的影响。方法下载并分析CGGA数据库RNAseq数据;收集人脑胶质瘤细胞系U251、U87和星形胶质细胞HA1800,用RT-qPCR法检测细胞系中的PDIA4表达水平;用siRNA转染U251细胞,沉默PDIA4的细胞株为si#768组和si#1829组,未转染为阴性对照组,转染siNC为空白对照组;用阿尔玛蓝(alarma blue)法、克隆形成实验检测转染细胞的增殖活力,流式细胞术检测细胞凋亡率;用蛋白免疫印记实验检测细胞中的cleaved-caspase3、cleaved-caspase 7蛋白表达水平。结果CGGA数据库中,PDIA4在不同级别胶质瘤中的表达水平不同(P<0.05),PDIA4高表达组较低表达组生存预后较差(Log-rank=264.32,P<0.001);与HA1800相比,PDIA4在U251和U87中相对表达量升高(F=58.44,P<0.05);与空白组、阴性对照组相比,si#768组和si#1829组增殖和克隆集落形成能力受到抑制(均P<0.01),凋亡率(17.67±0.11%、19.50±9.30%vs.3.72±7.11%、6.53±0.13%,F=51.06,P<0.05)明显升高;与阴性对照组相比,si#768组和si#1829组cleaved-caspase 7蛋白表达分别上调218.53%与169.68%,差异具有统计学意义(F=12.61,P<0.05)。结论沉默PDIA4基因可能通过激活caspase通路促进胶质瘤细胞凋亡。Objective To investigate the expression of protein disulfide isomerase family A member 4(PDIA4)in glioma and the effect on apoptosis.Methods The RNAseq data was downloaded from CGGA database.Glioblastoma cell lines(U251,U87)and human normal astrocyte cell line HA1800 were used in the study.The relative expression of PDIA4 in glioma cell lines was detected by RT⁃qPCR.U251 cells were transfected with siRNA,transfected with siPDIA4 as si#768 group and si#1829 group,untransfect⁃ed as negative control group,and transfected with siNC as blank control group.Then alarma blue assay and colony formation assay were used to examine the proliferation of U251 cells,the apoptosis rate was detected by flow cytometry,and Western blotting was used to detect the expression of cleaved⁃caspase3、cleaved⁃caspase7 protein.Results In CGGA database,the expression level of PDIA4 were different in gliomas of different grades(P<0.05),the five⁃year survival rates of the high PDIA4 expression group was lower than the low expression group,and Log⁃rank test(Log⁃rank=264.32,P<0.001).Compared with normal astrocyte cell line HA1800,the relative expression levels of PDIA4 was increased in U251 and U87(F=58.44,P<0.05).Compared with blank group and negative control group,proliferation and colony formation ability of si#768 group and si#1829 group were inhibited(P<0.01),and apoptosis rates(17.67±0.11%,19.50±2.30%vs.3.72±2.11%,6.53±0.13%,F=51.06,P<0.01)were significantly increased.Compared with the negative control group,the protein expression of cleaved⁃caspase⁃7 were increased up to 218.53%and 169.68%(F=12.61,P<0.05).Conclusion Knockdown of PDIA4 may promotes the apoptosis of glioma cells by activating caspase pathway.

关 键 词：蛋白二硫键异构酶家族成员4 胶质瘤 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]