Objective

To investigate how the interaction of CtBP2 with ZBTB18 affect glioblastoma (GBM).

Methods

Western blotting was performed to detect CtBP2 and ZBTB18 expression in GBM and normal brain tissues (NBT). U-87 MG cells were transfected with ZBTB18 CRISPR activation plasmid, CtBP2 shRNA with/without ZBTB18 shRNA. The biological characteristics were detected by EdU assay, MTT, Wound-healing, Transwell, TUNEL staining, and Flow cytometry. Furthermore, U-87 MG cells transfected with CtBP2 shRNA and/or ZBTB18 shRNA were injected into the flank region of mice and the tumor volume was measured. The mRNA and protein expression was quantified by qRT-PCR or Western blotting.

Results

GBM tissues exhibited increased CtBP2 expression and decreased ZBTB18 expression, which demonstrated a negative correlation in GBM tissues and showed the combined effect on prognosis. Based on immunoprecipitation and immunofluorescence, there was an interaction between CtBP2 and ZBTB18 in U-87 MG cells. CtBP2 shRNA counteracted the effect of ZBTB18 shRNA on inhibiting U-87 MG cell apoptosis, as well as promoting cell proliferation and viability with increased EMT, invasion and migration. Meanwhile, CtBP2 shRNA interact with ZBTB18 to block cells at phase G0/G1 and suppress SHH-GLI1 pathway. CtBP2 shRNA decreased tumor volume, increase ZBTB18 expression in tumor tissues, and inhibit SHH-GLI1 pathway in mice, which could be reversed by ZBTB18 shRNA.

Conclusion

CtBP2 elevation and ZBTB18 down-regulation were found in GBM, both of which were associated with prognosis of GBM patients. CtBP2 interacted with ZBTB18 to affect biological characteristics of GBM cells, and the tumor growth, which may be related to the SHH-GLI1 pathway.

中文翻译：

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CtBP2与ZBTB18相互作用以促进胶质母细胞瘤的恶性

目的

调查CtBP2与ZBTB18的相互作用如何影响胶质母细胞瘤（GBM）。

方法

进行了蛋白质印迹，以检测GBM和正常脑组织（NBT）中的CtBP2和ZBTB18表达。用具有/不具有ZBTB18 shRNA的ZBTB18 CRISPR激活质粒CtBP2 shRNA转染U-87 MG细胞。通过EdU分析，MTT，伤口愈合，Transwell，TUNEL染色和流式细胞仪检测生物学特性。此外，将用CtBP2 shRNA和/或ZBTB18 shRNA转染的U-87 MG细胞注射到小鼠的侧翼区域，并测量肿瘤体积。通过qRT-PCR或蛋白质印迹法定量mRNA和蛋白质表达。

结果

GBM组织表现出增加的CtBP2表达和ZBTB18表达降低，这表明GBM组织呈负相关，并显示出对预后的综合作用。基于免疫沉淀和免疫荧光，Ut-87 MG细胞中CtBP2和ZBTB18之间存在相互作用。CtBP2 shRNA抵消了ZBTB18 shRNA抑制U-87 MG细胞凋亡的作用，并通过增加EMT，侵袭和迁移来促进细胞增殖和存活。同时，CtBP2 shRNA与ZBTB18相互作用，阻断G0 / G1期的细胞并抑制SHH-GLI1途径。CtBP2 shRNA减少了肿瘤体积，增加了肿瘤组织中ZBTB18的表达，并抑制了小鼠的SHH-GLI1途径，这可能被ZBTB18 shRNA逆转。

结论

在GBM中发现CtBP2升高和ZBTB18下调，两者均与GBM患者的预后相关。CtBP2与ZBTB18相互作用，影响GBM细胞的生物学特性以及肿瘤的生长，这可能与SHH-GLI1途径有关。