载蟾毒灵Pluronic-PEI纳米微囊通过抑制miR-497介导的IGF1-R-PI3K-Akt通路抗大肠癌侵袭转移  

Bufalin-loaded Pluronic-PEI nanoparticles inhibits the invasion and metastasis of colorectal cancer via miRNA-497 mediated IGF1-R-PI3K-Akt signaling pathway

作　　者：胡明泰 胡强 殷佩浩[2] 段友容[3] 刘霖 欧阳常娟 谢东浩 曹雷 邱艳艳[2] 程韵枫[5] 王向东[5] 周明 邹瑜[2] 李荣鑫 

机构地区：[1]上海市徐汇区大华医院普外科,上海200237 [2]上海市普陀区中心医院肿瘤实验室,上海200062 [3]上海市肿瘤研究所,上海200031 [4]上海市徐汇区大华医院大中医科,上海200237 [5]复旦大学附属中山医院实验中心,上海200031

出　　处：《上海中医药杂志》2017年第10期89-96,共8页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市卫计委进一步加快中医药事业发展三年行动计划(2014年-2016年)资助项目(ZY3-CCCX-3-3054);中国临床医学诊疗技术创新发展专项基金项目(320.6750.14202)

摘　　要：目的探讨载蟾毒灵Pluronic-PEI纳米微囊通过抑制miR-497介导的IGF1-R-PI3K-Akt信号通路对大肠癌Lovo细胞体外及在裸鼠体内生长的影响及其作用机制。方法利用慢病毒miR-497转染大肠癌Lovo细胞,观察载蟾毒灵Pluronic-PEI纳米微囊对Lovo细胞体外增殖和侵袭转移能力的影响及miR-497介导的IGF1-R-PI3K-Akt信号通路相关基因和蛋白的调控作用。建立裸鼠原位移植瘤模型,8周后处死动物,观测对成瘤的影响。结果 (1)与对照组比较,载蟾毒灵的纳米微囊对过表达miR-497的Lovo细胞的抑制率最显著并呈时间-剂量依赖性;并使其凋亡率从(28.93±0.24)%提高至(75.20±0.29)%,明显高于相同浓度作用的Lovo细胞和Lovo NC细胞(P<0.01);划痕实验中过表达miR-497的Lovo细胞的迁移率从(0.083±0.019)(25 nmol/L)降低到(-0.242±0.035)(400 nmol/L),呈剂量依赖性;(2)Transwell结果提示载蟾毒灵纳米微囊在25 nmol/L就能够显著抑制过表达miR-497的Lovo细胞的侵袭作用;(3)免疫印迹分析提示载蟾毒灵的纳米微囊和LY 294002作用Lovo NC细胞和过表达miR-497的Lovo细胞,两种细胞的p-Akt Ser 473和p-Akt Thr 308蛋白表达明显下降,而pan-Akt变化不明显。(4)体内实验显示载蟾毒灵纳米微囊抑制瘤体转移优于蟾毒灵组。结论载蟾毒灵Pluronic-PEI纳米微囊可以通过抑制miR-497介导的IGF1-R-PI3K-Akt信号通路,在体外、体内都可发挥抗结肠癌侵袭和转移的作用。Objective To investigate the influence and mechanism of bufalin-baded Pluronicpolyetherimide （PEI） nanoparticles on the growth of eolorectal cancer Lovo cells in vitro and vivo via miR-497 mediated IGF1-R-PI3K-Akt signaling pathway. Methods The colorectal adenocarcinoma Lovo cells were transfected by lentivirus miR-497. The effects of bufalin-loaded Pluronic-PEI nanopartieles on proliferation, invasion and metastasis of Lovo cells in vitro were observed, as well as the regulation on the related gene and protein in miR-497 mediated IGF1-R-PBK-Akt signaling pathway. The orthotopic xenograft model of colorectal cancer was established in nude mice. The mice were sacrificed after 8 weeks, and the tumor formation was observed. Results ① Compared with the control group, the proliferation inhibition rate of bufalin-loaded Pluronic-PEI nanopartieles on miR-497 overexpressed Lovo ceils was the most significant and was increased in a time and dose dependent manner, and the apoptosis rate of miR-497 overexpressed Lovo cells was increased from （28.93 ± 0.24 ）% to （75.20 ± 0.29 ）% ,which was significantly higher than that of the Lovo ceils and Lovo NC ceils treated with the same concentration （P 〈 0.01 ）. In the scratch assay, the migration rate of miR- 497 overexpressed Lovo ceils was decreased from （0. 083 ± 0. 019 ） （25 nmol/L）to （- 0. 242 ± 0. 035 ） （400 nmol/L ） in a dose-dependent manner. ②The Transwell assay showed that bufalin-loeded Plurenic-PEI nanoparticles at concentration of 25 nmol/L could significantly inhibit the inyasion of miR-497 overexpressed Lovo cells. ③Western blot showed that the protein expressions of p-Akt Se1473 and p-Akt Thr308 in Lovo NC cells and miR-497 overexpressed Lovo cells were significantly decreased after the treatment with bufalin-loaded Pluronic-PEI nanoparticles and LY294002, but there was no obvious change on the expression of pan-Akt. ④In vivo experiments, the effects of bufalin-loaded Pluronic-PEI nanoparticles on inhibiting tumor metasta

关 键 词：蟾毒灵 纳米微囊 miR-497 IGF1-R-PI3K-Akt 大肠癌 

分 类 号：R285.5[医药卫生—中药学]