ABSTRACT
Objective: The most commonly reported primary brain tumor in adults is glioma. Choline kinase alpha (CHKA) has been proved to play important roles in glioma. However, the mechanism of CHKA involved remains unclear. Therefore, this study aims to explore the mechanism of CHKA in glioma development. Methods: Immunohistochemistry, qRT-PCR, and Western blot were used to detect the expression of CHKA. Flow cytometry, Cell Counting Kit-8 (CCK-8), transwell, and wound healing assays were performed to evaluate cell apoptosis, proliferation, invasion, and migration, respectively. RNA sequencing was used to explore the differentially expressed genes affected by CHKA. The enrichment analysis of gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) helped to detect the signaling pathways CHKA affected. Tumor-bearing mice were established and evaluated by TUNEL assay, Ki-67 immunohistochemistry. and hematoxylin and eosin staining. Results: CHKA increased in glioma tissues and promoted cell proliferation, invasion, and migration, while inhibiting the glioma cell apoptosis. It was also showed that CHKA promoted glioma development in vivo. GO and KEGG analysis indicated that PI3K/AKT was significantly enriched in CHKA knockdown U251 cells. And CHKA promoted glioma development by activating PI3K/AKT signaling pathway. Conclusions: The authors demonstrated that CHKA was significantly elevated in glioma tissues. Mechanism analysis indicated that CHKA could promote glioma development by activating PI3K/AKT signaling pathway, suggesting that CHKA is promising to be a biomarker and therapeutic strategy for prognostic prediction of patients with glioma.
ABSTRACT
Objective To detect the expression of choline kinase α (CHKA) in glioma and to further explore the effect of CHKA knockdown on the proliferation, invasion and migration of U87MG human glioma cells. Methods The mRNA expression of CHKA in high-grade gliomas and traumatic brain tissues was detected by real-time quantitative PCR. The expression of CHKA protein in high-grade gliomas and traumatic brain tissues was detected by Western blot analysis. The short hairpin RNA of CHKA (shCHKA) lentivirus and its control lentivirus (shNC) were constructed and used to infect U87MG glioma cells, which were then divided into the following three groups: shCHKA group, shNC group and blank control group. The proliferation of U87MG cells was examined by CCK-8 assay, the invasion ability of glioma cells was tested by TranswellTM invasion assay, and the migration ability of glioma cells was evaluated by scratch healing test. Results The CHKA mRNA and protein were highly expressed in glioma. Knockdown of CHKA gene inhibited the proliferation, invasion and migration of U87MG glioma cells. Conclusion The expression of CHKA in glioma tissue is significantly higher than that in the normal brain tissue, and knockdown of CHKA gene inhibits the proliferation, invasion and migration of glioma cells. It suggests that CHKA may be related to the occurrence and development of gliomas.