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Dynamic effect of bortezomib on nuclear factor-kappaB activity and gene expression in tumor cells
Myong-Hee Sung1, Lorena Bagain, Zhong Chen
1Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, Room B602, Bldg. 41, 41 Library Dr., Bethesda, MD 20892, USA. sungm@mail.nih.gov
Abstract:
Nuclear factor-kappaB (NF-kappaB) influences the initiation, progression, and maintenance of diverse cancer types. Despite current therapeutic efforts to block hyperactive NF-kappaB in cancer cells, the in vivo effects of a drug upon this complex pathway are unclear. We monitored NF-kappaB activity and a fast-expressing reporter level simultaneously in head and neck squamous carcinoma cells by quantitative live microscopy. The real-time single cell assay revealed the tumor necrosis factor-alpha-induced oscillation of NF-kappaB was echoed by equally dynamic reporter expression rate. Bortezomib is a proteasome inhibitor whose anticancer action is partly mediated through inhibition of NF-kappaB. When administered to preactivated cells, the drug gave rise to distinct inhibition dynamics, with discrete pulses of reporter induction remaining for hours. These findings suggest that, contrary to a simplistic presumption for a pathway "blockade," the network dynamics and the intracellular pharmacokinetics of the inhibitor must be critically evaluated in developing strategies for optimal intervention of oncogenic pathways.
Insights
Nuclear factor-kappaB (NF-kappaB) drives cancer progression. Real-time microscopy revealed bortezomib
Area of Science:
- Molecular Biology
- Cancer Research
- Pharmacology
Background:
- Nuclear factor-kappaB (NF-kappaB) is a critical regulator in cancer initiation, progression, and maintenance.
- Current therapeutic strategies targeting NF-kappaB often lack clarity regarding in vivo pathway dynamics.
- Understanding the real-time effects of drugs on complex signaling networks is crucial for effective cancer treatment.
Purpose of the Study:
- To simultaneously monitor NF-kappaB activity and reporter gene expression in real-time within head and neck squamous carcinoma cells.
- To investigate the dynamic effects of bortezomib, a proteasome inhibitor, on the NF-kappaB pathway in preactivated cancer cells.
- To evaluate the impact of intracellular pharmacokinetics and network dynamics on therapeutic intervention strategies for oncogenic pathways.
Main Methods:
- Quantitative live microscopy was employed to monitor NF-kappaB activity and reporter levels in single head and neck squamous carcinoma cells.
- Tumor necrosis factor-alpha (TNF-α) was used to induce NF-kappaB signaling.
- Bortezomib was administered to assess its inhibitory effects on the dynamic NF-kappaB pathway.
Main Results:
- Real-time single-cell analysis demonstrated that TNF-α-induced NF-kappaB oscillations were mirrored by dynamic reporter expression rates.
- Bortezomib treatment resulted in distinct inhibition dynamics, characterized by discrete pulses of reporter induction persisting for hours.
- These findings challenge the notion of a simple pathway blockade, highlighting complex drug-target interactions.
Conclusions:
- The study underscores the importance of considering network dynamics and intracellular pharmacokinetics when developing cancer therapies targeting the NF-kappaB pathway.
- A simplistic blockade approach may be insufficient; understanding dynamic responses is key for optimizing therapeutic strategies.
- Real-time single-cell assays provide valuable insights into drug effects on complex signaling networks in cancer.
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