Proteomic Profiling Reveals HECTD4-dependent Regulation of Protein Ubiquitination and Signaling Pathways in Prostate...

Area of Science:

• Oncology
• Molecular Biology
• Biochemistry

Background:

• Prostate cancer (PCa) is a significant male malignancy.
• Ubiquitination is a critical post-translational modification in cancer progression.
• HECTD4, a HECT-type E3 ubiquitin ligase, is implicated in cancer biology.

Purpose of the Study:

• Identify HECTD4 ubiquitination targets in PCa.
• Elucidate the role of HECTD4 in PCa molecular mechanisms.
• Explore HECTD4 as a potential therapeutic target for PCa.

Main Methods:

• HECTD4 knockdown in LNCaP, PC-3, and DU145 PCa cell lines.
• Semi-quantitative PCR and LC-MS/MS for protein expression and ubiquitination profiling.
• Gene ontology, pathway analysis, and proliferation assays.

Main Results:

• Identified over 3,300 differentially expressed proteins upon HECTD4 knockdown.
• NUSAP1, CDK6, and MED13L found to be ubiquitinated by HECTD4.
• Targets are linked to PI3K-AKT, Ras-MAPK, and mTOR pathways, immune infiltration, and drug response.

Conclusions:

• HECTD4 regulates protein stability and activation via ubiquitination in PCa.
• HECTD4 influences cell cycle, tumor suppression, and immune response.
• HECTD4 represents a promising therapeutic target for PCa treatment development.