Related Experiment Video
Updated: Jun 14, 2025
In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity
Published on: November 2, 2020
PCSK9 is a passenger gene in head and neck cancer with minimal pathological influence
Hyun-Ji Kim1, Dong-Guk Park1, Su-Jung Choi1
1Department of Oral Pathology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul 03080, Republic of Korea.
Objectives:
In this study, we aimed to explore the pathological effects of PCSK9 in head and neck cancer (HNC) by ablating its expression using the CRISPR-Cas9 knockout system.
Design:
To investigate the clinical importance of PCSK9 in HNC, in silico analysis was performed using datasets from the GEO database and the UALCAN database. To evaluate the role of PCSK9 in HNC pathogenesis, both CRISPR-Cas9 knockout system and pharmacological inhibition were employed to ablate PCSK9 expression in HNC cell lines. The impact of PCSK9 on cellular growth and proliferation was assessed using Cell Counting Kit-8, soft agar, and clonogenic assays in a 2D culture model, as well as a hanging drop spheroid formation assay with live/dead staining in a 3D culture model. The involvement of PCSK9 in apoptosis induction was evaluated by detecting c-PARP expression through Western blotting, measuring the sub-G1 population via cell cycle assay, and verifying the Annexin V-positive population. Finally, changes in metastatic profiles associated with fluctuations in PCSK9 expression were examined using wound healing and transwell migration/invasion assays.
Results:
In in silico analysis results, PCSK9 appeared to be related to the progression of HNC. However, experimental results demonstrated that PCSK9 plays a minimal role in cancer cell proliferation, anchorage-independent growth, colony formation capacity, in 2D cultures, as well as spheroidal growth in 3D cultures, and apoptosis induction. Furthermore, PCSK9 marginally influenced wound closure, metastatic potential, and invasive ability.
Conclusion:
Collectively, these data suggest that PCSK9 serves a neutral role in HNC, functioning as a passenger gene.
Insights
Proprotein convertase subtilisin/kexin type 9 (PCSK9) shows a neutral role in head and neck cancer (HNC) progression. Ablating PCSK9 expression minimally impacted HNC cell proliferation, apoptosis, and metastasis, suggesting it acts as a passenger gene.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Proprotein convertase subtilisin/kexin type 9 (PCSK9) is implicated in various cellular processes.
- Its role in head and neck cancer (HNC) pathogenesis remains largely unexplored.
- Understanding PCSK9's function in HNC is crucial for potential therapeutic strategies.
Purpose of the Study:
- To investigate the pathological effects and clinical importance of PCSK9 in head and neck cancer (HNC).
- To evaluate the role of PCSK9 in HNC cell proliferation, apoptosis, and metastasis.
- To determine if PCSK9 acts as a driver or passenger gene in HNC development.
Main Methods:
- In silico analysis using GEO and UALCAN databases to assess PCSK9's clinical relevance in HNC.
- CRISPR-Cas9 gene knockout and pharmacological inhibition to ablate PCSK9 expression in HNC cell lines.
- In vitro assays (2D and 3D cultures) to evaluate proliferation, anchorage-independent growth, apoptosis, migration, and invasion.
Main Results:
- In silico analysis suggested a correlation between PCSK9 and HNC progression.
- Experimental ablation of PCSK9 had minimal impact on HNC cell proliferation, colony formation, and spheroid growth.
- PCSK9 demonstrated negligible effects on apoptosis induction, wound healing, and cell migration/invasion.
Conclusions:
- PCSK9 appears to play a neutral role in head and neck cancer.
- The findings suggest PCSK9 functions as a passenger gene in HNC.
- Further research may be needed to fully elucidate PCSK9's role in cancer biology.
More Related Videos
Related Concept Videos
01:05Single Nucleotide Polymorphisms-SNPs
02:26Comparing Copy Number Variations and SNPs
Copy number variations or CNVs are the structural variations that cover more than 1kb of DNA sequence. The single nucleotide polymorphism (SNP), on the other hand, is a single nucleotide change or a point mutation that is found in more than 1%...