基于3D细胞培养模型的新型纳米载药系统抗胆囊癌活性的效果及作用机制  

Anti-gallbladder carcinoma activity of a novel nano-drug delivery system based on 3D cell culture model

作　　者：江翰[1] 王子豪[1] 李永盛[1] 边睿[1] 施伟斌[1] 

机构地区：[1]上海交通大学医学院附属新华医院普外科,上海200092

出　　处：《临床肝胆病杂志》2014年第3期264-268,共5页Journal of Clinical Hepatology

基　　金：上海市科委纳米专项项目(11nm0503700)

摘　　要：目的通过建立胆囊癌3D细胞培养模型,研究新型纳米载药系统对胆囊癌细胞的杀伤效果及作用机制。方法构建胆囊癌细胞GBC-SD的2D/3D培养模型,利用MTT、HE染色、流式细胞仪(FCM)、Hoechst/PI凋亡检测等方法探索叶酸修饰的聚己内酯基药物P(CL-co-OPD)纳米载药系统CP24对正常细胞的毒性以及载药后对胆囊癌细胞的抑制率计量资料比较采用t检验。结果 HE染色显示纳米载药系统CP24对肝肾细胞未见明显细胞毒性。FCM法显示CP24对胆囊癌细胞无明显杀伤作用。在氟尿嘧啶(5-Fu)浓度为100μg/ml时,5-Fu-CP24组以及单纯5-Fu组在3D细胞模型下对肿瘤的杀伤和抑制效果均较2D细胞模型下减低。无论在2D/3D细胞模型下,5-Fu-CP24组对肿瘤细胞的杀伤效果均显著强于单纯5-Fu组(P<0.05),通过Hoechst/PI凋亡检测前者引起细胞的死亡以凋亡为主,后者以坏死为主。结论 3D细胞培养可以更加真实的反映体内细胞之间的微环境,细胞间基质的影响可能使细胞对药物的敏感性降低。携5-Fu的纳米载体比原药5-Fu能引起更多的细胞死亡,这种可降解纳米载药系统具有良好的载药性、缓释性及生物安全性,其对于胆囊癌的化疗具有广阔的应用前景。Objective To establish a three -dimensional （3D） cell culture model for gallbladder carcinoma （GC） cells and to investigate the killing effect of a novel nano - drug delivery system on GC cells and its action mechanism. Methods Two - dimensional （2D） and 3D cell culture models for GC GBC - SD cells were established, and the normal cell toxicity of nano - drug delivery system （ CP24, folate recep tor - mediated targeting poly （ epsilon - caprolactone） ） and the inhabitation rate of GC cells exposed to 5 - fluorouracil （5 - FU ） carried by CP24 were determined by MTI＂ assay, HE staining, flow cytometry （ FCM）, and Hoechst/PI staining. Results CP24 showed no significant cytotoxicity in liver and kidney cells, according to the result of HE staining. CP24 showed no significant killing effect on GC cells, according to the result of FCM. Given a 5 - FU concentration of 100 μg/ml, the killing and inhibitory effects of both 5 - FU - CP2d and 5 - FU on GBC - SD cells were lower when using the 3D cell culture model than when using the 2D cell culture model; whether using a 2D or 3D cell culture model, 5 - FU - CP24 had a significantly stronger killing effect on GBC - SD cells than 5 - FU （ P 〈 0.05 ）. 5 - FU - CP24 mainly led to apoptosis in GBC - SD cells, while 5 - FU mainly led to necrosis, as shown by Hoechst/PI staining. Conclusion 3 D cell culture can reflect the intracellular microenvironment in vivo more accurately. Intercellular matrix may affect the cellular sensitivity to drugs. 5 - FU can kill more GC cells when carried by a nano - drug delivery system. This degradable nano - drug delivery system has good drug delivery per formance, sustained drug release, and high biosafety and holds promise for application in chemotherapy for GC.

关 键 词：胆囊肿瘤 细胞培养技术 钠米医学 抗肿瘤联合化疗方案 

分 类 号：R735.8[医药卫生—肿瘤]