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Published on: August 25, 2021
Tumour suppressive effects of WEE1 gene silencing in neuroblastomas
Ahmad Hosseini Tashnizi, Mansooreh Jaberipour, Mahboobeh Razmkhah
1Department of Immunology, Immunotherapy Laboratory, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Aim Of Study:
WEE1, a member of serine/threonine protein kinase family is the master inhibitor of cyclin-dependent kinase 1 in cell cycle. Over-expression of WEE1 in glioblastomas (GBMs) and some other cancers has been shown. Here, we investigated the expression of WEE1 in 13 brain samples from GBM patients and two GBM cell lines. Further to that, we asked whether if knocking down WEE1 expression in the cell lines change tumor cells' reaction.
Materials And Methods:
All brain tumor samples were collected after confirmed pathological diagnosis. Western blotting was used to screen the expression of WEE1 and a panel of tumor markers. As a model of WEE1 gene silencing with small hairpin RNA (shRNA) technology in GBMs, A172, and U373GM cell lines were transfected with four WEE1 specific shRNAs. The growth characteristics of the cells and the expression of a panel of downstream genes were investigated after gene suppression.
Results:
All GBMs and both cell lines over-expressed WEE1. Transduction of the cell lines with different shRNAs suppressed WEE1 expression with different extent and pooling of four shRNAs together resulted in additive effect. Suppression of WEE1 not only repressed cellular growth but also changed the profile of gene expression of the cells. Quantitative real-time polymerase chain reaction showed also reduced expression of genes such as hypoxia-inducible factor-1, B-cell lymphoma-2, vascular endothelial growth factor, and p53 with crucial roles in tumor survival and invasiveness.
Conclusion:
These results highlight the key role of WEE1 suppression to combat GBMs. Moreover, it showed beneficial possibilities of WEE1 suppression with different anticancer approaches for neurological malignancies.
Insights
WEE1 kinase is overexpressed in glioblastomas (GBMs). Suppressing WEE1 kinase in GBM cells reduced tumor growth and altered expression of key survival genes, suggesting WEE1 inhibition as a potential GBM therapy.
Area of Science:
- Molecular Biology
- Oncology
- Cell Cycle Regulation
Background:
- WEE1 kinase is a master inhibitor of cyclin-dependent kinase 1, crucial for cell cycle progression.
- Overexpression of WEE1 has been observed in glioblastomas (GBMs) and other cancers.
- Investigating WEE1's role in GBMs is critical for understanding tumor development and identifying therapeutic targets.
Purpose of the Study:
- To investigate the expression levels of WEE1 in human glioblastoma samples and cell lines.
- To determine the effect of WEE1 gene knockdown on GBM cell growth and gene expression.
- To explore the potential of WEE1 suppression as a therapeutic strategy for glioblastomas.
Main Methods:
- Analysis of WEE1 expression in 13 GBM patient brain samples and two GBM cell lines (A172, U373GM) using Western blotting.
- Gene silencing of WEE1 in GBM cell lines utilizing small hairpin RNA (shRNA) technology.
- Assessment of cellular growth characteristics and downstream gene expression profiles (including HIF-1, BCL-2, VEGF, p53) post-WEE1 suppression via quantitative real-time PCR.
Main Results:
- WEE1 was overexpressed in all analyzed GBM samples and cell lines.
- shRNA-mediated WEE1 knockdown effectively suppressed GBM cell proliferation.
- Suppression of WEE1 led to reduced expression of crucial tumor survival and invasiveness genes, including hypoxia-inducible factor-1, B-cell lymphoma-2, vascular endothelial growth factor, and p53.
Conclusions:
- WEE1 plays a significant role in glioblastoma progression and survival.
- Targeted suppression of WEE1 demonstrates a promising therapeutic avenue for combating GBMs.
- WEE1 inhibition may synergize with other anticancer strategies for treating neurological malignancies.
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