shRNA沉默TXNDC12抑制脑胶质瘤细胞的增殖和迁移  

Silencing TXNDC12 by shRNA inhibits the proliferation and migration of glioma cells

作　　者：张路瑶 袁红平[2] 王晓旋 李晓全[2] 李泽艳 王雨晴 来明名 马雪艳 ZHANG Lu-yao;YUAN Hong-ping;WANG Xiao-xuan;LI Xiao-quan;LI Ze-yan;WANG Yu-qing;LAI Ming-ming;MA Xue-yan(Department of Biochemistry and Molecular Biology,School of Basic Medical,Dali University,Dali 671000,China;Department of Neurosurgery,the Third Affiliated Hospital of Kunming Medical University,Kunming 650000,China;Department of Pathology,the Third People’s Hospital of Jinan,Jinan 250000,China;Department of Blood Transfusion,the First Affiliated Hospital of Dali University,Dali 671000,China)

机构地区：[1]大理大学基础医学院生物化学与分子生物学教研室,大理671000 [2]昆明医科大学第三附属医院神经外科,昆明650000 [3]山东省济南市第三人民医院病理科,济南250000 [4]大理大学第一附属医院输血科,大理671000

出　　处：《临床与实验病理学杂志》2023年第5期519-524,共6页Chinese Journal of Clinical and Experimental Pathology

基　　金：国家自然科学基金(82160504);云南省科技厅科技计划(202101AU070136)。

摘　　要：目的探讨TXNDC12(thioredoxin domain-containing 12)在脑胶质瘤发生中的作用机制。方法采用生物信息学分析TXNDC12基因在脑胶质瘤中mRNA的表达及其与预后的相关性,并通过Western blot检测TXNDC12在脑胶质瘤组织和正常脑组织中蛋白表达;构建沉默TXNDC12基因的shRNA慢病毒表达质粒,建立沉默TXNDC12的U251稳转细胞株。采用qRT-PCR和Western blot实验分别检测稳转细胞株中TXNDC12的mRNA及蛋白表达;用CCK-8实验、Transwell和划痕实验检测沉默TXNDC12对U251细胞增殖和迁移能力的影响。结果生物学信息分析显示TXNDC12在脑胶质瘤组织(1.885±0.902)中的蛋白表达比正常脑组织(1.0±0.545,P<0.05)高,且与不良预后相关(高表达组51.5%vs低表达组70.7%,P<0.0001);沉默TXNDC12后U251细胞增殖能力和迁移能力明显下降(P<0.05)。结论TXNDC12在脑胶质瘤中可能发挥促癌基因的作用,其高表达可能促进脑胶质瘤细胞的增殖和迁移,有望成为脑胶质瘤患者治疗及预后的重要生物学标志物。Purpose To investigate the potential role of the stable U251 cell lines(TXNDC12)in gliomas.Methods The mRNA expression differences of TXNDC12 in giloma tissues and normal tissues were analyzed based on bioinformatics methods,and the protein expression of TXNDC12 was detected in glioma tissue and normal brain tissue by Western blot.The shRNA silencing expression plasmid of TXNDC12 gene was constructed and identified,and then U251 giloma cells were infected with shTXNDC12 lentivirals,the stable U251 cell lines(shTXNDC12)with knockdown TXNDC12 were established,qRT-PCR and Western blot were performed to detect the mRNA and protein level of TXNDC12.CCK-8 assay,Transwell assay and scratch assay were used to detect the effects of TXNDC12 knockdown on proliferation and migration of U251 cells.Results The protein expression of TXNDC12 in glioma tissues(1.856±0.880)was higher than that in adjacent normal brain tissues(1.135±0.619,P<0.05),and it was associated with poor prognosis(high expression group 51.5%vs low expression group 70.7%,P<0.0001).The proliferation and migration abilities of U251 cells were significantly decreased(P<0.05)after TXNDC12 was silenced.Conclusion TXNDC12 may play an oncogenic role in glioma,and its high expression may promote the proliferation and migration of glioma cells,which is expected to become an important potential biomarker for the treatment and prognosis of patients with glioma.

关 键 词：脑胶质瘤 TXNDC12 U251细胞 增殖 迁移 

分 类 号：R739.41[医药卫生—肿瘤]