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Effects of NFkappaB decoy oligonucleotides released from biodegradable polymer microparticles on a glioblastoma cell
Jagjit S Gill1, Xun Zhu, Michael J Moore
1Molecular Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA.
Abstract:
The objectives of this study were to investigate a nuclear factor-kappa B (NFkappaB) decoy oligonucleotide (ODN) strategy on the inhibition of glioblastoma (GBM) cell line growth and to evaluate a poly(DL-lactic-co-glycolic acid) (PLGA) microparticle delivery system for the NFKB decoy ODNs in vitro. We have demonstrated that NFkappaB activation is important in regulating GBM cell line growth. Aberrant nuclear expression of NFkappaB was found in a panel of GBM cell lines, while untransformed glial cells did not display NFkappaB activity. Nuclear translocation of NFkappaB was inhibited by using a 'decoy" ODN strategy. NFkappaB decoy ODNs designed to inhibit NFkappaB resulted in a significant reduction in cell number (up to 45%) compared to control cultures after 2 days. The reduction in cell number correlated with a decrease in cyclin D1 protein expression and a commensurate decrease in Cdk-4 activity. These results provide evidence suggesting that NFkappaB mediates cell cycle progression and demonstrates a mechanism linking increased NFkappaB activity with GBM cell growth and cell cycle disregulation. Decoy ODNs were encapsulated at a yield of 66% in PLGA microparticles and released in a controlled manner in phosphate buffered saline for up to 28 days. Approximately 83% of entrapped ODNs were released by day 28. During 3 days of GBM cell line culture, the released decoy ODNs retained their biologic activity and led to significantly reduced cell number as compared to control cultures. These findings offer a potential therapeutic strategy in the control of human GBM cell line growth in vitro and suggest that PLGA microparticles may be appropriate as delivery vehicles for the "decoy" ODN strategy.
Insights
A nuclear factor-kappa B (NF-kappaB) decoy oligonucleotide strategy significantly reduced glioblastoma cell growth by inhibiting cell cycle progression. Poly(DL-lactic-co-glycolic acid) microparticles effectively delivered these decoy oligonucleotides, showing therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Nuclear factor-kappa B (NF-kappaB) is aberrantly expressed in glioblastoma (GBM) cell lines and drives their growth.
- NF-kappaB activation is crucial for regulating GBM cell proliferation and cell cycle progression.
- Untransformed glial cells do not exhibit the same level of NF-kappaB activity as GBM cells.
Purpose of the Study:
- To investigate the efficacy of a nuclear factor-kappa B (NF-kappaB) decoy oligonucleotide (ODN) strategy in inhibiting glioblastoma (GBM) cell growth.
- To evaluate the use of poly(DL-lactic-co-glycolic acid) (PLGA) microparticles as a delivery system for NF-kappaB decoy ODNs in vitro.
- To elucidate the mechanism by which NF-kappaB influences GBM cell cycle regulation.
Main Methods:
- Utilized NF-kappaB decoy oligonucleotides designed to inhibit nuclear translocation of NF-kappaB.
- Encapsulated decoy ODNs within PLGA microparticles for controlled release.
- Assessed cell number, cyclin D1 protein expression, and Cdk-4 activity in GBM cell lines.
- Monitored ODN release kinetics from PLGA microparticles in phosphate-buffered saline.
Main Results:
- NF-kappaB decoy ODNs significantly reduced GBM cell number by up to 45% compared to controls.
- The reduction in cell number correlated with decreased cyclin D1 protein expression and Cdk-4 activity.
- PLGA microparticles demonstrated a 66% yield for ODN encapsulation and sustained release of active ODNs for up to 28 days.
- Released decoy ODNs from PLGA microparticles retained biological activity, significantly reducing GBM cell growth in vitro.
Conclusions:
- NF-kappaB plays a critical role in mediating GBM cell cycle progression and growth.
- The NF-kappaB decoy ODN strategy presents a potential therapeutic approach for controlling human GBM cell growth.
- PLGA microparticles are suitable delivery vehicles for implementing the decoy ODN strategy against GBM in vitro.