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Published on: July 17, 2020
TMEM16F Regulates Baseline Phosphatidylserine Exposure and Cell Viability in Human Embryonic Kidney Cells
Unlabelled:
Background / Aims: TMEM16F is a transmembrane protein from a conserved family of Ca2+-activated proteins, which is highly expressed in several tissues. TMEM16F confers phospholipid scramblase activity and Ca2+-activated electrolyte channel activity. Potentially thereby, TMEM16F is involved in cell cycle control and apoptotic signaling. The present study evaluated the role of TMEM16F on cell proliferation and viability in Human Embryonic Kidney cells.
Methods:
An inducible knockdown of TMEM16F was generated and markers of apoptosis and proliferation were assessed via flow cytometry, western blotting and MTT uptake assay under different conditions.
Results:
TMEM16F knockdown resulted in attenuated growth of HEK293 cells. This observation correlated with an increased phosphatidylserine exposure and a decreased fraction of viable cells. Interestingly, the cells were not sensitized to Staurosporine- induced cell death. Western blot analyses displayed a parallel activation of pro- and antiapoptotic signaling pathways: Caspase 3 cleavage and Cyclin D1 abundance were simultaneously increased. Furthermore, knockdown of TMEM16F led to activation of AKT signaling.
Conclusion:
TMEM16F modifies viability of Human Embryonic Kidney cells via its function as a phospholipid scramblase and activation of AKT signaling pathways.
Insights
Transmembrane protein 16F (TMEM16F) regulates human embryonic kidney cell viability and proliferation. Its knockdown reduces cell growth and viability, impacting phospholipid scramblase and AKT signaling pathways.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- TMEM16F is a Ca2+-activated transmembrane protein family member.
- It exhibits phospholipid scramblase and Ca2+-activated electrolyte channel activity.
- TMEM16F is implicated in cell cycle control and apoptotic signaling.
Purpose of the Study:
- To investigate the role of TMEM16F in cell proliferation and viability.
- To assess TMEM16F's impact on Human Embryonic Kidney (HEK293) cells.
Main Methods:
- Generated inducible TMEM16F knockdown in HEK293 cells.
- Assessed apoptosis and proliferation markers using flow cytometry and Western blotting.
- Utilized MTT uptake assay to evaluate cell viability.
Main Results:
- TMEM16F knockdown attenuated HEK293 cell growth and viability.
- Phosphatidylserine exposure increased, while viable cell fraction decreased.
- Observed simultaneous activation of pro- and anti-apoptotic pathways (Caspase 3, Cyclin D1) and AKT signaling.
Conclusions:
- TMEM16F influences Human Embryonic Kidney cell viability.
- Its function as a phospholipid scramblase and AKT pathway activation are key mechanisms.
