Gill Jagjit S, Zhu Xun, Moore Michael J, Lu Lichun, Yaszemski Michael J, Windebank Anthony J
Molecular Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA.
Biomaterials. 2002 Jul;23(13):2773-81. doi: 10.1016/s0142-9612(02)00013-3.
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.
本研究的目的是探讨核因子-κB（NFκB）诱饵寡核苷酸（ODN）策略对胶质母细胞瘤（GBM）细胞系生长的抑制作用，并在体外评估聚（DL-乳酸-共-乙醇酸）（PLGA）微粒对NFκB诱饵ODN的递送系统。我们已经证明NFκB激活在调节GBM细胞系生长中很重要。在一组GBM细胞系中发现NFκB的异常核表达，而未转化的神经胶质细胞未显示NFκB活性。使用“诱饵”ODN策略可抑制NFκB的核转位。与对照培养物相比，设计用于抑制NFκB的NFκB诱饵ODN在2天后导致细胞数量显著减少（高达45%）。细胞数量的减少与细胞周期蛋白D1蛋白表达的降低以及Cdk-4活性的相应降低相关。这些结果提供了证据，表明NFκB介导细胞周期进程，并证明了一种将NFκB活性增加与GBM细胞生长和细胞周期失调联系起来的机制。诱饵ODN以66%的产率封装在PLGA微粒中，并在磷酸盐缓冲盐水中以可控方式释放长达28天。到第28天，约83%的包封ODN被释放。在GBM细胞系培养的3天中，释放的诱饵ODN保留了其生物活性，与对照培养物相比导致细胞数量显著减少。这些发现为体外控制人GBM细胞系生长提供了一种潜在的治疗策略，并表明PLGA微粒可能适合作为“诱饵”ODN策略的递送载体。
