摘要
[目的]探究ZBTB7A对脑胶质瘤的影响及其改善脑胶质瘤裸鼠生存时间的作用机制。[方法]将U87细胞(每组接种2 mL、5×10^(5)个/mL)随机分为3组:U87-WT组(常规培养)、U87-NC组(500μL/孔感染100μg/mL包装空载质粒的慢病毒)、U87-ZBTB7A组(500μL/孔感染100μg/mL包装过表达ZBTB7A重组质粒的慢病毒)。构建稳定过表达ZBTB7A的U87细胞系(加200μL/孔、1μg/mL嘌呤霉素筛选)。用qRT-PCR、蛋白免疫印迹法检测各个细胞系中ZBTB7A的表达量;平板克隆、细胞划痕、Transwell实验检测ZBTB7A对U87细胞的增殖、迁移和侵袭能力影响;流式细胞术检测ZBTB7A对U87细胞凋亡的影响;蛋白免疫印迹检测ZBTB7A、TRAIL-R2表达水平的变化;裸鼠原位接种脑胶质瘤,评估ZBTB7A对裸鼠生存的影响。[结果]过表达ZBTB7A显著抑制了U87细胞的增殖、侵袭和迁移能力(P<0.05)。过表达ZBTB7A促进了U87细胞的细胞凋亡。Western blot检测结果显示,ZBTB7A的过表达激活了TRAIL-R2的表达(P<0.05)。接种了ZBTB7A过表达的U87细胞的裸鼠具有更长的生存时间。[结论]过表达ZBTB7A通过上调TRAIL-R2进而抑制了脑胶质瘤细胞增殖、侵袭和迁移,促其凋亡,延长接种脑胶质瘤细胞裸鼠的生存时间。
[Objective]To explore the effects of ZBTB7A on glioma and the mechanism of ZBTB7A improving the survival time of brain glioma in nude mice.[Method]The U87 cells(with density of 5×10^(5) cells/mL and culture volume of 2 mL)were randomly divided into the U87-WT group(routine culture),the U87-NC group(infected with 100μg/mL packaging empty plasmid lentivirus,with a dosage of 500μL per well),and the U87-ZBTB7A group(infected with 100μg/mL packaged overexpressed ZBTB7A recombinant plasmid lentivirus,with a dosage of 500μL per well).A stable U87 cell line overexpress-ing ZBTB7A was constructed(adding 200μL of 1μg/mL puromycin solution to each well for screening).The expression of ZBTB7A was detected by qRT-PCR and Western blot.The effects of ZBTB7A on proliferation,migration and invasion of U87 cells were detected by plate cloning assay,wond-healing assay and Transwell assay,respectively.The effect of ZBTB7A on ap-optosis of U87 cells was detected by flow cytometry.The expression levels of ZBTB7A and TRAIL-R2 were detected by West-ern blot.To evaluate the effect of ZBTB7A on survival of nude mice,glioma cells were inoculated in situ.[Result]Overexpres-sion of ZBTB7A significantly inhibited the proliferation,migration and invasion of U87 cells(P<0.05).Overexpression of ZBTB7A promoted apoptosis of U87 cells.Western blot results showed that overexpression of ZBTB7A activated the expression of TRAIL-R2(P<0.05).Nude mice inoculated with ZBTB7A overexpressed U87 cells survived longer than other nude mice in other groups.[Conclusion]Overexpression of ZBTB7A inhibits the proliferation,invasion and migration of glioma cells by upregulating TRAIL-R2 and promotes their apoptosis,prolonging the survival time of nude mice inoculated with glioma cells.
作者
董海军
郭俊霞
梁盛
朱亮亮
杨峰
DONG Haijun;GUO Junxia;LIANG Sheng;ZHU Liangliang;YANG Feng(Department of Neurosurgery,Handan Hospital of Integrated Traditional Chinese and Western Medicine,Handan 056001;Department of Hematology,Handan First Hospital,Handan 056000;Department of Pharmacy,Cangzhou Central Hospital,Cangzhou 061000;Department of Neurosurgery,Cangzhou Central Hospital,Cangzhou 061000;Department of Neurosurgery,Affiliated Hospital of North China University of Science and Technology,Tangshan 063000,China)
出处
《生物技术》
2025年第6期726-732,共7页
Biotechnology
基金
河北省医学科学研究课题计划资助(20220345)。
