人胰腺癌PANC-1细胞裸鼠移植瘤模型建立及其用于观察载siRNA纳米微粒体内效应的研究  

Establishment of human pancreatic tumor xenograft mouse model for evaluating tumor-homing and gene-silencing effects of siRNA-loading nanoparticles

作　　者：曾林涓[1] 李景果[1] 张秋波[1] 钱辰琛[1] 林忠[1] 陈茵婷[1] 黄开红[1] 

机构地区：[1]中山大学附属第五医院肿瘤内科,广东珠海519000

出　　处：《中国病理生理杂志》2014年第3期572-576,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81072045;No.81302140);广东省产学研资助项目(No.2009B090300277);珠海市科技计划(No.2013D0401990026)

摘　　要：目的:探讨建立人胰腺癌PANC-1裸鼠移植瘤模型的最佳实验方法,并应用该模型进行载基因纳米微粒体内效应的研究。方法:将不同数量PANC-1细胞悬液接种于BALB/c(nu/nu)小鼠右侧背部皮下,当肿瘤体积达100 mm3时尾静脉注射siRNACY5.5纳米复合物进行活体荧光成像。此外,于尾静脉注射负载siRNAKras纳米复合物,蛋白印迹及免疫组织化学染色法观察肿瘤组织Kras蛋白表达水平。结果:1×107cells/300μL接种成瘤率达100%,成瘤时间<2周。荧光呈像及组织学检查显示载siRNA纳米微粒可靶向聚集在肿瘤组织发挥体内基因沉默效应。结论:本研究报道的人胰腺癌裸鼠移植瘤模型建立方法成瘤率达100%,成瘤时间短,是研究药物示踪和观察疗效的理想模型。AIM： To establish an effective and rapid method to develop transplanted subcutaneous pancreatic carcinoma by inducing PANC-1 cells into nude mice, and then use this mouse model to evaluate the tumor-homing and gene-silencing effects of siRNA-loading nanoparticles in vivo. METHODS ： Different numbers of PANC-1 ceils in 100 uL or 300 uL PBS were inoculated subcutaneously into the fight flank of BALB/c （nu/nu） mice. When the tumor volume reached 100 mm3, siRNAcv5.5 nanoparticles were injected through the mouse tail vein to perform in vivo imaging assay. Be sides, the mice were randomly divided into 3 treatment groups treated with PBS, scrambled control RNA nanoparticles and siKras nanoparticles, respectively. The protein expression of Kras was detected by Western blotting and immunohistocbemi cal staining. RESULTS： After inoculated with 1 x 107 PANC-1 cells in 300 uL PBS, all mice developed tumors within 2 weeks. The in vivo results showed that siRNA-loading nanoparticles accumulated in the tumor tissues and exerted gene si lencing effect. CONCLUSION： In the present study, an effective and rapid method was established for PANC-1 cells to induce transplanted subcutaneous pancreatic carcinoma in nude mice within 2 weeks, which is suitable for in vivo imaging and treatment evaluations as a reproducible and reliable way for the further experiments.

关 键 词：胰腺肿瘤 模型 动物 纳米微粒 RNA干扰 

分 类 号：R363[医药卫生—病理学]