PO.ET02.08 · 实验与分子治疗
具有优化胆固醇浓度和源自模型低级别浆液性卵巢癌细胞系脂质提取物的纳米脂质体的细胞摄取
Cellular uptake of nanoliposomes with optimized cholesterol concentration and lipid extract derived from a model low grade serous ovarian carcinoma cell line
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：低级别浆液性卵巢癌（LGSOC）是上皮性卵巢癌的一种罕见亚型，占所有病例的不到10%，通常在较年轻女性中诊断。该疾病的特征是由MAPK通路的激活突变（包括KRAS、BRAF和NRAS）驱动的肿瘤生长。尽管进展缓慢，LGSOC由于对常规化疗的反应有限，仍然难以治疗。本研究的目的是从一个模型LGSOC细胞系中分离细胞脂质提取物（LE）材料，并利用新获得的LE材料开发一种相对靶向特异的纳米脂质体系统。早期制剂和体外研究包括对胆固醇和LE含量的优化。
方法：培养并扩增低级别浆液性卵巢癌细胞系（PM-LGSOC-01，Cytion）于补充10% FBS的EMEM生长培养基中。当细胞达到约90%汇合度时，从PM-LGSOC-01中提取PM-LGSOC-01脂质提取物（LE）材料。脂质提取按先前所述方法进行（Alharbi & Campbell，AAPS Open，编号：5(2018)）。纳米脂质体制剂由不同比例的DOPC、胆固醇和LE组成，采用薄膜法通过Buchi R-80旋转蒸发仪形成。制剂中加入DPPE-Rhodamine用于细胞研究。超声处理后，使用ZetaPals测定粒径和zeta电位值。荧光检测使用荧光酶标仪进行。
结果：制备了三种制剂：(1) DOPC（100%），(2) DOPC/Chol（95/5），(3) DOPC/Chol（90/10），平均粒径分别为221±3 nm、250±5 nm和139±0.3 nm。初步结果表明，在纳米脂质体制剂中额外加入胆固醇增加了靶（PM-LGSOC-01）细胞对其的摄取。关于PM-LGSOC-01-LE对靶细胞摄取纳米脂质体的影响的研究目前正在进行中。
结论：迄今为止，已研究了胆固醇对LGSOC细胞摄取纳米脂质体的影响。未来的研究将评估优化的胆固醇和LE含量对靶向低级别浆液性卵巢癌的影响。
查看英文原文 English abstract
Introduction: Low-grade serous ovarian carcinoma (LGSOC) represents a rare subtype of epithelial ovarian cancer, accounting for less than 10% of all cases and typically diagnosed in younger women. The disease is characterized by tumor growth driven by activating mutations in the MAPK pathway, including KRAS , BRAF , and NRAS . Despite its slow progression, LGSOC remains challenging to treat due to limited responsiveness to conventional chemotherapy. The objective of this study was to isolate cellular lipid extract (LE) material from a model LGSOC cell line, and to utilize the newly acquired LE material to develop a relatively target specific nanoliposomal system. Early formulation and in vitro studies include optimization for cholesterol and LE content.
Methods: Low-grade serous ovarian carcinoma cell line (PM-LGSOC-01, Cytion) was cultured and expanded in EMEM growth medium supplemented with 10% FBS. The PM-LGSOC-01-lipid extract (LE) material was extracted from PM-LGSOC-01 when the cells reached ~90% confluency. Lipid extraction was performed as described previously (Alharbi & Campbell, AAPS Open, number: 5(2018)). Nanoliposomal preparations consisted of DOPC, cholesterol, and LE at various ratios and wereformed by thin film method using a Buchi R-80 rotary evaporator. DPPE-Rhodamine was included in the preparations for cellular studies. Following sonication, particle size and zeta potential values weredetermined using ZetaPals. Fluorescence detection was performed using a fluorescence microplate reader.
Results: Three preparations (1) DOPC (100%), (2) DOPC/Chol (95/5), and (3) DOPC/Chol (90/10) wereprepared with an average particle size of 221 ± 3 nm, 250 ± 5 nm, and 139 ± 0.3 nm, respectively. Preliminary results suggest that the additional inclusion of cholesterol in the nanoliposomal preparations increased their uptake by the target (PM-LGSOC-01) cells. Studies investigating the influence of PM-LGSOC-01-LE on the uptake of nanoliposomes by the target cells are currently underway.
Conclusion: To date, the influence of cholesterol on the uptake of nanoliposomes by LGSOC cells has been investigated. Future studies will evaluate the effect of optimized cholesterol and LE contenton targeting low-grade serous ovarian carcinoma.
利益披露 Disclosure
S. Orlando, None..
K. Panchal, None..
H. Patel, None..
R. B. Campbell, None.