Knockdown of CREB3 inhibits proliferation, migration, and invasion of glioma cells by decreasing phosphorylated FAK (Tyr397) expression

doi:10.3877/cma.j.issn.1674-0785.2023.02.016

Abstract

Abstract:

Objective

To investigate the relationship between cAMP response binding protein 3 (CREB3) gene and the survival prognosis of glioma patients, and its functional role and mechanism in malignant biological behaviors of glioma cells, such as proliferation, invasion, and metastasis, so as to provide a theoretical basis for finding effective molecular therapeutic targets for glioma.

Methods

The data on the mRNA expression level of CREB3 and survival rate were obtained from the Chinese Glioma Genome Atlas (CCGA) database. Immunocytochemistry (IHC) was performed to verify the expression of CREB3 and phosphorylated focal adhesion kinase 397 (p-FAK) protein in different grades of human glioma tissues. Lentiviruses were used to construct CREB3 knockdown models in the glioma cell lines U87 and U251. Western blot (WB) was used to verify the expression of p-FAK and related proteins in the FAK signaling pathway, as well as related apoptotic proteins.Cell Counting Kit-8 was used to calculate the optical density (OD) of cells at 450 nm, plate cloning assay was used to verify the proliferation ability of cells after knockdown of CREB3, and cell scratch and Transwell assays were used to verify the migration and invasion ability of knockdown cell lines.

Results

According to the CGGA database, CREB3 expression was found to be up-regulated in high-grade gliomas, indicating a poor prognosis (P＜0.05). IHC showed that CREB3 expression increased with increasing glioma grade, while p-FAK (397) was significantly expressed in grade IV gliomas. Knockdown of CREB3 decreased the expression of p-FAK (397), increased the expression of the pro-apoptotic protein BCL2-associated X (BAX), decreased the expression of the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) (P＜0.05), and inhibited the proliferation, migration, and invasion of glioma cells in vitro (P＜0.05).

Conclusion

CREB3 expression is increased in high-grade glioma and is closely related to the survival prognosis of patients. Knockdown of CREB3 downregulates the phosphorylation of FAK in glioma cells, which in turn reduces the proliferation, invasion, and migration of glioma cells.

Key words: Glioma, U87-MG, U251, Cell invasion, Cell migration, Cell proliferation

Yiwei Zhang, Yaxin Hu, Liangzhao Chu, Zhao Yan, Qian Zeng, Qian Pu. Knockdown of CREB3 inhibits proliferation, migration, and invasion of glioma cells by decreasing phosphorylated FAK (Tyr397) expression[J]. Chinese Journal of Clinicians(Electronic Edition), 2023, 17(02): 202-209.

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URL: https://zhlcyszz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-0785.2023.02.016

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