Page 7 - 南京医科大学学报自然科学版

第43卷第3期 南京医科大学学报（自然科学版）
2023年3月 Journal of Nanjing Medical University（Natural Sciences） ·297 ·

·基础研究·

ZBTB3表达对胶质母细胞瘤细胞增殖和克隆形成的调控

王伟民 ，赵晨卉 ，倪思琦 ，何庆玲 ，阮玉婷 ，吴宁霞 ，张 婧 ，王迎伟 ，邱 文 1*
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南京医科大学免疫学系，江苏 南京 211166；南京医科大学第一附属医院肿瘤科，江苏 南京 210029；南京医科大学第
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一临床医学院临床医学系，江苏 南京 211166
［摘 要］ 目的：检查人胶质母细胞瘤（glioblastoma，GBM）组织和细胞系中ZBTB3的表达，并探讨ZBTB3对GBM细胞增殖和
克隆形成的影响及其调控机制。方法：通过GEPIA2数据库分析GBM患者肿瘤组织中ZBTB3的表达情况。RT⁃PCR、qPCR和
Western blot 检测 GBM 细胞系（U251、U373、U87）中 ZBTB3 的 mRNA 和蛋白表达水平，筛选出 ZBTB3 表达最高的 U87 细胞。
CCK⁃8和克隆形成实验检测沉默ZBTB3基因对U87细胞增殖和克隆形成的影响。用p38MAPK、AMPK、Akt1抑制剂处理U87
细胞后，Western blot 检测p38MAPK、AMPK、Akt1的磷酸化水平，RT⁃PCR、qPCR和Western blot 测定ZBTB3的mRNA和蛋白表
达水平，CCK⁃8 和克隆形成实验检测细胞增殖和克隆形成。结果：GBM 患者肿瘤组织中 ZBTB3 的表达显著高于正常组织。
U251、U373和U87细胞中均可见ZBTB3的表达，其中U87细胞表达最高。沉默ZBTB3基因能明显抑制U87细胞的增殖和克隆
形成。抑制AMPK既能显著降低U87细胞ZBTB3的表达水平，又可明显减弱U87细胞增殖和克隆形成。结论：GBM组织和细
胞系中ZBTB3的表达显著上调，GBM细胞中AMPK活化并上调ZBTB3基因的表达，促进GBM细胞的增殖和克隆形成。
［关键词］ 胶质母细胞瘤；ZBTB3；AMPK；增殖；克隆形成
［中图分类号］ R739.41 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）03⁃297⁃07
doi：10.7655/NYDXBNS20230301

Regulation of ZBTB3 expression on proliferation and clone formation of glioblastoma cells

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WANG Weimin ，ZHAO Chenhui ，Ni Siqi ，HE Qingling ，RUAN Yuting ，WU Ningxia ，ZHANG Jing ，WANG
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Yingwei ，QIU Wen 1*
1 Department of Immunology，Nanjing Medical University，Nanjing 211166；Department of Oncology，the First
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Affiliated Hospital of Nanjing Medical University，Nanjing 210029；Clinical Medical Department，the First Clinical
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Medical College，Nanjing Medical University，Nanjing 211166，China
［Abstract］ Objective：This study aims to examine the expression of ZBTB3 in human glioblastoma（GBM）tissues and cell lines，and
to explore the effects of ZBTB3 on the proliferation and clonal formation of GBM cells and their regulatory mechanism. Methods：The
expression of ZBTB3 in tumor tissues of GBM patients was analyzed by GEPIA2 database. The mRNA and protein expression levels of
ZBTB3 in GBM cell lines（U251，U373，U87）were detected by RT⁃PCR，qPCR and Western blot，and U87 cell line was identified with
the highest expression of ZBTB3. CCK ⁃ 8 and clonal formation assay were used to examine the effects of silencing ZBTB3 on the
proliferation and clonal formation of U87 cells. U87 cells were treated with p38MAPK，AMPK and Akt1 inhibitors，and the
phosphorylation levels of p38MAPK，AMPK and Akt1 were detected by Western blot. ZBTB3 mRNA and protein levels were detected
by RT⁃PCR，qPCR and Western blot. Cell proliferation and clone formation were examined by CCK⁃ 8 and clone formation assay.
Results：The expression of ZBTB3 in tumor tissues of GBM patients was significantly higher than that in normal tissues. The
expression levels of ZBTB3 in U251，U373 and U87 cell lines were examined，and the highest expression in U87 cells was observed.
Silencing ZBTB3 markedly inhibited the proliferation and clonal formation of U87 cells. AMPK inhibition could not only obviously
reduce the expression level of ZBTB3 in U87 cells，but also markedly attenuate the proliferation and clonal formation of U87 cells.
Conclusion：The expression of ZBTB3 is obviously increased in GBM tissues and cells，and the AMPK ⁃ up ⁃ regulated ZBTB3

［基金项目］ 国家自然科学基金（81971468）；江苏省高等学校大学生创新训练计划项目（202010312039Y）
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通信作者（Corresponding author），E⁃mail：qiuwen@njmu.edu.cn；wangyw1508@njmu.edu.cn