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Expression of STK11 gene and its promoter activity in MCF control and cancer cells
Asma Alkaf1, Abdulaziz Al-Jafari1, Tanveer A Wani2
1Department of Biochemistry, College of Science, King Saud University, Riyadh, 11495 Kingdom of Saudi Arabia.
Abstract:
Serine/threonine kinase gene (STK11) is identified as tumor suppressor gene whose mutation can lead to Peutz-Jeghers syndrome (PJS). STK11 is emerging as a multifunctional protein, it activates 14 different AMP-activated protein kinase (AMPK) family members, important in the regulation of cell polarity, cell cycle arrest, energy and hemostasis. Present study was designed to evaluate STK11 mRNA expression in MCF-7 cancer and MCF-10 normal breast cells lines. mRNA expression was studied by real-time PCR. Further, human STK11 promoter construct was fused to a luciferase reporter and transfected into both MCF-7 and MCF-10 cells to identify the promoter activity in these cells. STK11 mRNA was found significantly higher in MCF-7 compared to MCF-10 cells (p value < 0.0005) indicating its role in the onset of breast cancer. Interestingly, it was found that the promoter activity of STK11 gene in MCF-7 cells was also significantly higher when compared to MCF-10 cells (p value < 0.005). Positive correlation was observed in promoter activity and gene expression (p = 0.048, r2 = 0.587). This study for the first time relates the altered STK11 gene expression in breast cancer cells with altered promoter activity. The present finding may shed light on the new therapeutic approaches against breast cancer by targeting gene or its promoter.
Insights
The serine/threonine kinase 11 (STK11) gene shows higher expression and promoter activity in breast cancer cells (MCF-7) compared to normal cells (MCF-10). This suggests STK11
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- The serine/threonine kinase 11 (STK11) gene functions as a tumor suppressor, and its mutations are linked to Peutz-Jeghers syndrome (PJS).
- STK11 is a multifunctional protein that regulates cell polarity, cell cycle arrest, energy, and hemostasis through AMP-activated protein kinase (AMPK) family members.
Purpose of the Study:
- To evaluate STK11 mRNA expression in MCF-7 (breast cancer) and MCF-10 (normal breast) cell lines.
- To assess the promoter activity of the STK11 gene in both cancer and normal breast cell lines.
- To investigate the correlation between STK11 gene expression and its promoter activity in breast cancer.
Main Methods:
- Real-time PCR was used to quantify STK11 mRNA expression.
- Luciferase reporter assays were employed to measure STK11 promoter activity after transfecting constructs into MCF-7 and MCF-10 cells.
Main Results:
- STK11 mRNA expression was significantly higher in MCF-7 cells compared to MCF-10 cells (p < 0.0005).
- STK11 promoter activity was also significantly elevated in MCF-7 cells relative to MCF-10 cells (p < 0.005).
- A positive correlation was found between STK11 promoter activity and gene expression (p = 0.048, r² = 0.587).
Conclusions:
- This study demonstrates for the first time that altered STK11 gene expression in breast cancer cells is associated with altered promoter activity.
- The findings suggest a potential role for STK11 and its promoter in breast cancer development.
- Targeting the STK11 gene or its promoter may offer novel therapeutic strategies for breast cancer.
