PO.ET02.08 · 实验与分子治疗
SU-CCS-1脂质提取物修饰纳米脂质体及模型透明细胞肉瘤靶细胞系的特征性生长谱
SU-CCS-1 lipid extract-modified nanoliposomes and characteristic growth profile of a model clear cell sarcoma target cell line
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：肉瘤约占美国成人癌症的1%和儿童癌症的15%。常规治疗策略在疾病早期有一定效果，但一旦发生转移，治疗成功率显著下降。纳米医学提供了一种替代方法，以提高现有治疗疗效并延长生存期。脂质体制剂的优化对于确保最具选择性的靶向至关重要。本研究的目的是使用人类透明细胞肉瘤的细胞模型在体外开发一种新型纳米脂质体药物递送系统。
方法：选择SU-CCS-1细胞系作为透明细胞肉瘤的细胞模型并用于药物选择性研究。SU-CCS-1细胞在体外培养和扩增以用于细胞提取。细胞脂质提取物（LE）材料源自透明细胞肉瘤靶（SU-CCS-1）细胞系。纳米脂质体针对LE和胆固醇含量进行了优化。使用DPPE-Rhodamine（作为荧光指示剂）进行细胞摄取研究，并使用荧光酶标仪评估相对荧光强度值。评估的第I阶段包括对细胞的表征以及由DOPC和胆固醇组成、含0、5和10 mol%胆固醇含量的纳米脂质体制剂的细胞摄取特性。为表征细胞生长特征，用从成熟混合细胞群体中获取的100%贴壁细胞或100%悬浮细胞分别制备了单独的培养瓶。在第0天和第5天测定贴壁细胞和悬浮细胞的百分比。
结果：对SU-CCS-1靶细胞系生长谱特征的评估揭示了悬浮细胞随时间显著转变为贴壁细胞。100%贴壁细胞的培养瓶在5天后仍保持72%贴壁，而100%悬浮细胞的培养瓶在5天后转变为59%贴壁。含0、5和10 mol%胆固醇的纳米脂质体的粒径分别为283 nm、139.9 nm和247 nm。此外，与0% LE对照制剂（不含SU-CCS-1-LE）相比，纳入10 mol% SU-CCS-1-LE增加了纳米脂质体的摄取。
结论：初步发现表明，DOPC:胆固醇比例影响SU-CCS-1细胞对纳米脂质体的摄取。在纳米脂质体中纳入SU-CCS-1-LE增强了其靶向效率。正在进行的研究将探讨优化的胆固醇和SU-CCS-1-LE含量的双重掺入对透明细胞肉瘤靶向的影响。
查看英文原文 English abstract
Background: Sarcomas account for approximately 1% of adult cancers and 15% of childhood cancers in the United States. Conventional treatment strategies are somewhat effective during early disease, but once metastasized, treatment success significantly declines. Nanomedicine presents an alternative approach to increase current treatment efficacy and prolong survival. Optimization of liposomal preparations is essential to ensure the most selective targeting. The objective of the study was to develop a novel nanoliposomal drug delivery system using a cellular model of human clear cell sarcoma in vitro.
Methods: The SU-CCS-1 cell line was selected as the cellular model for clear cell sarcoma and used for the drug selectivity studies. SU-CCS-1 cells were cultured and expanded in vitro for cellular extraction purposes. Cellular lipid extract (LE) material was derived from a clear cell sarcoma target (SU-CCS-1) cell line. Nanoliposomes were optimized for LE and cholesterol content. DPPE-Rhodamine (used as a fluorescence indicator) for cellular uptake studies, and a fluorescence microplate reader was used to assess relative fluorescence intensity values. Phase I of evaluation included characterization of cells and cellular uptake properties of nanoliposomal preparations of DOPC and cholesterol consisting of 0, 5, and 10 mol% cholesterol content. To characterize the cellular growth characteristics, separate flasks were prepared with either 100% adherent or 100% suspension cells acquired from a mature mixed cell population. The percentage of adherent and suspension cells was determined on Day 0 and Day 5.
Results: An evaluation of growth profile characteristics of the SU-CCS-1 target cell line revealed a substantial transformation of suspension cells to adherent cells over time. While the flask of 100% adherent cells remained 72% adherent after 5 days, the flask of 100% suspension cells transformed to become 59% adherent after 5 days. The particle size for nanoliposomes containing 0, 5, and 10 mol% cholesterol was 283 nm, 139.9 nm, and 247 nm, respectively. Additionally, the inclusion of 10 mol% SU-CCS-1-LE increased uptake of nanoliposomes compared to 0% LE control preparations (without SU-CCS-1-LE).
Conclusion: Preliminary findings suggest that the DOPC:cholesterol ratio affects nanoliposome uptake by SU-CCS-1 cells. The inclusion of SU-CCS-1-LE in nanoliposomes enhances their targeting efficiency. Ongoing studies will investigate the effect of dual incorporation of optimized cholesterol and SU-CCS-1-LE content on the targeting of clear cell sarcoma.
利益披露 Disclosure
A. Silva, None..
A. Chan, None..
S. Banoud, None..
C. Bouchard, None.