Expression of PEA3 and EPHA2 in glioblastoma and role in the Wnt/β-catenin pathway

doi:10.3877/cma.j.issn.2095-123X.2024.02.002

Abstract

Abstract:

Objective

To explore the expression of polyomavirus enhancer activator 3 (PEA3) and erythropoietin-producting hepatocellular receptor 2 (EPHA2) in glioblastoma and their expression in Wnt/β-catenin pathway.

Methods

A gene transfection model of U87 glioma cells was constructed, and the cell lines were divided into 5 groups: blank group, PEA3 interference group, PEA3 interference empty group, EPHA2 interference group, and EPHA2 interference empty group. The protein expression levels of EPHA2 and PEA3 in U87 cells were detected through Western blotting experiments; The proliferation rate was detected by CCK-8 test, and the the expression levels of T cytokines factor-4 (TCF-4) and lymph enhancer factor 1 (LEF1) genes, the downstream genes of Wnt/β-catenin pathway, were detected by quantitative real time polymerase chain reaction (qRT-PCR) assay.

Results

The Western blotting experiment showed that compared with the blank group, the expression level of PEA3 was reduced in the PEA3 interference group and EPHA2 interference group, while the expression level of EPHA2 was reduced in the EPHA2 interference group, and the differences were statistically significant (P<0.05); There was no statistically significant difference in the expression levels of Wnt1 and β-catenin proteins among 5 groups (P>0.05). The CCK-8 experiment showed that compared with the blank group, the cell proliferation rates of the PEA3 interference group and the EPHA2 interference group were significantly reduced, and the differences were statistically significant (P<0.05). The qRT-PCR results showed that the expression levels of TCF-4 and LEF1 genes decreased in the PEA3 interference group and EPHA2 interference group, and the differences were statistically significant (P<0.05).

Conclusion

Interference with PEA3 and EPHA2 genes can reduce the proliferation ability of glioblastoma; whether PEA3 and EPHA2 promote the proliferative capacity of glioblastoma through the Wnt/β-catenin pathway deserves further investigation.

Key words: Polyomavirus enhancer activator 3, Erythropoietin-producting hepatocellular receptor 2, Glioblastoma, Wnt/β-catenin pathway

Tuoheti Maimaitiyiming, Ye Liu, Cheng Zhang, Yasen Abudukadier, Feng Gao, Jichao Wang, Yonggang Wu. Expression of PEA3 and EPHA2 in glioblastoma and role in the Wnt/β-catenin pathway[J]. Chinese Journal of Brain Diseases and Rehabilitation(Electronic Edition), 2024, 14(02): 73-79.

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Figures/Tables 7

Tab.1 Primer sequence

基因	序列
β-actin	F-CCCATCTATGAGGGTTACGC
β-actin	R-TTTAATGTCACGCACGATTTC
PEA3	F-GATGAAAGCCGGATACTTGGAC
PEA3	R-TTCGCGCAAGCTCCCATTT
EPHA2	F-TGGCTCACACACCCGTATG
EPHA2	R-GTCGCCAGACATCACGTTG
TCF-4	F-GCAGCAAGTCCGAGAAAGGA
TCF-4	R-GTCTCCCATTCCAGGGTGTG
LEF1	F-CTTCCATGTCCAGGTTTTCCC
LEF1	R-GGCTCCTGCTCCTTTCTCTG

Fig.1 Plasmid transfection map of U87 cells (×400)

Fig.2 Detection of transfection efficiency of PEA3 and EPHA2 genes

Fig.3 Western blotting results of EPHA2, PEA3, Wnt1, β-catenin of 5 groups

Fig.4 Expression levels of EPHA2, PEA3, Wnt1, β-catenin protein in 5 groups

Fig.5 Comparison of cell proliferation rates among 5 groups

Fig.6 Comparison of expression levels of TCF-4 and LEF1 genes in 5 groups

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