ODF3B affects the proliferation and apoptosis of glioma via the JAK/STAT pathway

Abstract

The pathogenesis of glioma has remained unclear. In this study, it was found that high expression of the outer dense fibers of sperm tail 3B (ODF3B) in gliomas was positively correlated with the grade of glioma. The higher the grade, the worse the prognosis. ODF3B is closely related to the growth and apoptosis of glioma. In terms of mechanism, ODF3B was found to affect the proliferation and apoptosis of glioma through the JAK1 and JAK2/STAT3 pathways. ODF3B was also found to affect the growth and apoptosis of glioma in vivo. We conclude that ODF3B affects glioma proliferation and apoptosis via the JAK/STAT pathway and is a potential therapeutic target.

Keywords: Glioma; JAK/STAT; ODF3B; apoptosis; proliferation.

Figure 1

ODF3B is highly expressed in glioma and positively correlated with grade. A. Relative expression of ODF3B in common tumors in the TCGA database. B-D. The expression of ODF3B in glioma samples collected from the TCGA database, CGGA database and the Affiliated Hospital of Southwest Medical University relative to that in normal tissues. E-G. Relationship between ODF3B expression and tumor grade in the TCGA database, CGGA database and samples collected from the Affiliated Hospital of Southwest Medical University. H, I. The expression of ODF3B in glioma cell lines was verified by qPCR and Western blotting. J. The expression of ODF3B in glioma tissues and normal tissues was verified by immunohistochemistry (*P < 0.05, **P < 0.01, ***P < 0.001).

Figure 2

Relationship between ODF3B expression and clinicopathological features. A, B. The expression of ODF3B in the TCGA database and CGGA database was correlated with age. C, D. ODF3B expression was correlated with IDH mutation status in the TCGA and CGGA databases. E. The relationship between 1p19q deletion and ODF3B expression in the TCGA database. F. The relationship between histology and ODF3B expression in the TCGA database (*P < 0.05, **P < 0.01, ***P < 0.001).

Figure 3

High expression of ODF3B predicts poor prognosis. A. ODF3B in the CGGA was correlated with overall survival (OS). B. ODF3B in the TCGA database was correlated with OS. C. ODF3B was associated with disease-specific survival (DSS) in the TCGA database. D. ODF3B was associated with progression-free interval (PFI) in the TCGA database.

Figure 4

The decrease in ODF3B affects the proliferation and apoptosis of glioma. A, B. Validation of low knockdown efficiency of glioma cell lines using qPCR and Western blotting after lentivirus knockdown. C, D. Proliferation ability of glioma cells after ODF3B knockdown. E, F. Colony formation experiment after ODF3B knockdown. G. Changes in apoptosis after ODF3B knockdown. H, I. Expression changes of apoptosis-related proteins after ODF3B knockdown (*P < 0.05, **P < 0.01, ***P < 0.001).

Figure 5

ODF3B affects the proliferation and apoptosis of glioma through the JAK/STAT pathway. A. In TCGA database, KEGG analysis of relevant pathways. B. In the TCGA database, gene set enrichment analysis (GSEA) was used, and JAK/STAT was a significantly enriched tumor-related pathway in patients with high ODF3B expression. C, D. After ODF3B knockdown, Western blotting (WB) verified the changes in JAK/STAT pathway proteins. E. Changes in apoptotic protein and the JAK/STAT pathway after Colivelin TFA was added to lentivirus knockdown detected by WB. F. WB was used to detect lentivirus knockdown and Colivelin TFA was added. CCK8 was used to detect cell proliferation. G, H. Change in apoptosis ability detected by WB after adding Colivelin TFA after lentivirus knockdown (*P < 0.05, **P < 0.01, ***P < 0.001).

Figure 6

Down-regulation of ODF3B can affect the development of glioma in vivo. A. Comparison of tumor size between the two groups after in situ tumor formation in nude mice. B. Changes in tumor Ki-67 and cleaved-caspase 3 in nude mice after knocking down ODF3B. C. Comparison of survival time between the two groups of nude mice after knocking down ODF3B (*P < 0.05, **P < 0.01, ***P < 0.001).