Sensitising PDAC to Gemcitabine by Suppressing NF-κB Pathway and Enhancing Apoptosis

Area of Science:

• Oncology
• Molecular Biology
• Drug Discovery

Background:

• Pancreatic ductal adenocarcinoma (PDAC) demonstrates poor response to gemcitabine due to chemoresistance.
• There is a critical need for agents that enhance gemcitabine efficacy without increasing toxicity.

Purpose of the Study:

• To characterize a small molecule, B12, as a potential gemcitabine sensitizer for PDAC.
• To evaluate B12's ability to overcome gemcitabine resistance in PDAC cell models.

Main Methods:

• MTT assays to determine IC50 and dose-modifying factor (DMF).
• Colony formation, wound scratch, JC-1, and Annexin V/PI assays for phenotypic and apoptotic analysis.
• mRNA-seq for transcriptomic profiling and NF-κB pathway analysis via p65 fractionation and RT-qPCR.

Main Results:

• B12 alone showed minimal cytotoxicity but reduced gemcitabine's IC50 by 1.39-fold in PDAC cells.
• Co-treatment with B12 and gemcitabine enhanced apoptosis, inhibited proliferation, and reduced colony formation.
• Transcriptomic analysis revealed B12 downregulated genes involved in growth and survival, associated with reduced NF-κB signaling.

Conclusions:

• B12 effectively sensitizes PDAC cells to gemcitabine with low intrinsic toxicity.
• The sensitizing effect is linked to increased apoptosis and suppressed NF-κB signaling.
• Further validation is needed to confirm causality and lack of sensitization in normal cells.