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Tumour suppressive effects of WEE1 gene silencing in breast cancer cells
Naghmeh Ghiasi1, Mojtaba Habibagahi, Rozita Rosli
1Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Background:
WEE1 is a G2/M checkpoint regulator protein. Various studies have indicated that WEE1 could be a good target for cancer therapy. The main aim of this study was to asssess the tumor suppressive potential of WEE1 silencing in two different breast cancer cell lines, MCF7 which carries the wild-type p53 and MDA-MB468 which contains a mutant type.
Materials And Methods:
After WEE1 knockdown with specific shRNAs downstream effects on cell viability and cell cycle progression were determined using MTT and flow cytometry analyses, respectively. Real-time PCR and Western blotting were conducted to assess the effect of WEE1 inhibition on the expression of apoptotic (p53) and anti-apoptotic (Bcl2) factors and also a growth marker (VEGF).
Results:
The results showed that WEE1 inhibition could cause a significant decrease in the viability of both MCF7 and MDA-MB-468 breast cancer cell lines by more than 50%. Interestingly, DNA content assays showed a significant increase in apoptotic cells following WEE1 silencing. WEE1 inhibition also induced up- regulation of the apoptotic marker, p53, in breast cancer cells. A significant decrease in the expression of VEGF and Bcl-2 was observed following WEE1 inhibition in both cell lines.
Conclusions:
In concordance with previous studies, our data showed that WEE1 inhibition could induce G2 arrest abrogation and consequent cell death in breast cancer cells. Moreover, in this study, the observed interactions between the pro- and anti-apoptotic proteins and decrease in the angiogenesis marker expression confirm the susceptibility to apoptosis and validate the tumor suppressive effect of WEE1 inhibition in breast cancer cells. Interestingly, the levels of the sensitivity to WEE1 silencing in breast cancer cells, MCF7 and MDA-MB468, seem to be in concordance with the level of p53 expression.
Insights
WEE1 inhibition significantly reduced breast cancer cell viability and increased apoptosis by altering p53, Bcl2, and VEGF levels. This validates WEE1 as a tumor suppressor target in breast cancer therapy.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- WEE1 kinase is a G2/M checkpoint regulator with potential as a cancer therapeutic target.
- This study investigates the tumor suppressive role of WEE1 inhibition in breast cancer.
Purpose of the Study:
- To assess the impact of WEE1 silencing on cell viability and cell cycle progression in MCF7 and MDA-MB-468 breast cancer cell lines.
- To evaluate the effects of WEE1 inhibition on key apoptotic and growth-related markers.
Main Methods:
- WEE1 knockdown was achieved using specific shRNAs.
- Cell viability was assessed by MTT assay, and cell cycle progression by flow cytometry.
- Real-time PCR and Western blotting were used to analyze p53, Bcl2, and VEGF expression levels.
Main Results:
- WEE1 inhibition decreased viability in both cell lines by over 50%.
- Significant increases in apoptosis and p53 (apoptotic marker) expression were observed.
- Downregulation of Bcl2 (anti-apoptotic marker) and VEGF (angiogenesis marker) was noted.
Conclusions:
- WEE1 inhibition abrogates the G2/M checkpoint, leading to cell death in breast cancer cells.
- The observed changes in apoptotic and angiogenesis markers confirm WEE1 inhibition's tumor suppressive effects.
- Breast cancer cell sensitivity to WEE1 silencing correlates with p53 expression levels.
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