HeLa Cervical Cancer Cells Are Maintained by Nephronophthisis 3-Associated Primary Cilium Formation via ROS-Induced ERK and HIF-1α Activation under Serum-Deprived Normoxic Condition
Abstract
:1. Introduction
2. Results
2.1. Serum-Deficiency Contributes to PC Formation and the Expression of NPHP3 and HIF-1α
2.2. HIF-1α Regulates NPHP3 Expression and PC Formation
2.3. SD-Induced ROS Are Associated with NPHP3 Expression and HIF-1α-Mediated PC Formation
2.4. ROS-Increased ERK Is Related to PC Formation and NPHP3 Transcription
2.5. PC Formation Influences NPHP3 Expression and Cell Growth in SD Conditions
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Plasmids and siRNAs
4.3. Cell Culture
4.4. Cytotoxicity Assay
4.5. Hypoxia Treatment
4.6. ROS Measurement
4.7. Detection of Primary Cilia
4.8. Transfection of Nucleic Acids
4.9. Gaussia Luciferase Assay for Promoter Activity
4.10. Chromatin Immunoprecipitation (ChIP) Assay
4.11. Reverse Transcription Polymerase Chain Reaction (RT-PCR)
4.12. Real-Time Reverse Transcription (Q-PCR) Analysis
4.13. Western Blotting
4.14. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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Lee, J.-W.; Cho, J.-Y.; Thuy, P.X.; Moon, E.-Y. HeLa Cervical Cancer Cells Are Maintained by Nephronophthisis 3-Associated Primary Cilium Formation via ROS-Induced ERK and HIF-1α Activation under Serum-Deprived Normoxic Condition. Int. J. Mol. Sci. 2022, 23, 14500. https://doi.org/10.3390/ijms232314500
Lee J-W, Cho J-Y, Thuy PX, Moon E-Y. HeLa Cervical Cancer Cells Are Maintained by Nephronophthisis 3-Associated Primary Cilium Formation via ROS-Induced ERK and HIF-1α Activation under Serum-Deprived Normoxic Condition. International Journal of Molecular Sciences. 2022; 23(23):14500. https://doi.org/10.3390/ijms232314500
Chicago/Turabian StyleLee, Jae-Wook, Jun-Yeong Cho, Pham Xuan Thuy, and Eun-Yi Moon. 2022. "HeLa Cervical Cancer Cells Are Maintained by Nephronophthisis 3-Associated Primary Cilium Formation via ROS-Induced ERK and HIF-1α Activation under Serum-Deprived Normoxic Condition" International Journal of Molecular Sciences 23, no. 23: 14500. https://doi.org/10.3390/ijms232314500