纳米颗粒递送siRNA联合声动力疗法协同治疗结肠癌的实验研究  

Nanoparticle-delivered siRNA combined with sonodynamic therapy for colon cancer

作　　者：李琢[1] 陈真 王华强 肖志刚[1] LI Zhuo;CHEN Zhen;WANG Huaqiang;XIAO Zhigang(Department of Colorectal and Anal Surgery,Hunan Provincial People's Hospital/the First Affiliated Hospital of Hunan Normal University,Changsha 410005,China;Hunan Provincial Key Laboratory of Micro&Nano Materials Interface Science,College of Chemistry and Chemical Engineering,Central South University,Changsha 410083,China)

机构地区：[1]湖南师范大学附属第一医院/湖南省人民医院结直肠肛门外科,湖南长沙410005 [2]中南大学化学化工学院湖南省微纳米材料界面科学重点实验室,湖南长沙410083

出　　处：《中国普通外科杂志》2024年第10期1654-1666,共13页China Journal of General Surgery

基　　金：湖南省肛肠疾病质量控制中心基金资助项目(20220307-1006)。

摘　　要：背景与目的:声动力疗法(SDT)是一种新兴的肿瘤治疗方法,主要利用低强度超声(US)激活声敏剂,促使其产生细胞毒性的活性氧(ROS)杀伤细胞。本研究尝试开发一种包含声敏剂与肿瘤相关基因RRS1小干扰RNA(siRRS1)的新型纳米颗粒(NPs),并观察其对结肠癌的抗癌作用,以期为结肠癌的治疗提供新的思路。方法:在TCGA与GEO数据库下载RRS1 RNA-Seq数据,收集湖南省人民医院结直肠肛门外科80例临床结肠癌患者的临床病理资料,分析RRS1在结肠癌中的表达及其与结肠癌预后的关系。观察结肠癌细胞转染siRRS1后的增殖与凋亡情况。将声敏剂四(4-羧基苯基)卟啉(TCPP)、基因载体DSPE-PEI、siRRS1组装成TCPP@DSPE-PEI-siRRS1 NPs(T@D-siRRS1 NPs),在体外对T@D-siRRS1 NPs进行结构表征,以及其细胞摄取与ROS生成能力检查后,进行体外细胞实验与在体荷瘤鼠实验观察T@D-siRRS1NPs对结肠癌的抗癌作用。结果:数据库分析与临床标本检测均显示,RRS1在结肠癌组织中表达高于癌旁正常组织;并且高表达与预后不良相关(HR=1.88,95%CI=1.27~2.79,P=0.002);单因素与多因素分析显示,RRS1 mRNA表达是结肠癌患者总生存期的独立危险因素(HR=1.354,95%CI=1.447~2.215,P=0.031)。敲低RRS1后,结肠癌细胞增殖能力明显减弱、凋亡明显增加(均P<0.05)。结构表征结果显示颗粒粒径均一,稳定性好;T@D-siRRS1 NPs通过细胞内吞作用进入细胞后,TCPP在超声的辐照下可以产生单线态氧(1O2)。随后的体内外实验结果均显示,转染携带siRRS1或TCPP(T@D-对照siRNA、T@D-siRRS1)+US的NPs后,结肠癌细胞的生长均有不同程度的抑制,其中T@D-siRRS1 NPs+US的作用最强(均P<0.05)。此外,T@D-siRRS1 NPs+US治疗小鼠的重要器官无明显损伤。结论:T@D-siRRS1 NPs+US集SDT与基因治疗于一体,对结肠癌细胞可产生有效的协同杀伤作用,且T@D-siRRS1 NPs具有良好的安全性与生物兼容性,具有临床应用的潜在可能。Background and Aims:Sonodynamic therapy(SDT)is an emerging tumor treatment method that primarily utilizes low-intensity ultrasound(US)to activate sonosensitizers,leading to the production of cytotoxic reactive oxygen species(ROS)that kill cells.This study was conducted with an attempt to develop a novel nanoparticle(NPs)formulation containing a sonosensitizer and the small interfering RNA(siRRS1)of tumor-related gene RRS1 to observe its anticancer effects on colon cancer,to provide new strategies for colon cancer treatment.Methods:RRS1 RNA-Seq data were downloaded from the TCGA and GEO databases,and clinical pathological data were collected from 80 patients with colon cancer at Hunan Provincial People's Hospital.The expression of RRS1 in colon cancer and its relationship with prognosis were analyzed.The proliferation and apoptosis of colon cancer cells transfected with siRRS1 were observed.The sonosensitizer tetra(4-carboxyphenyl)porphyrin(TCPP),gene carrier DSPE-PEI,and siRRS1 were assembled into TCPP@DSPE-PEI-siRRS1 NPs(T@D-siRRS1 NPs).After structural characterization and examination of cellular uptake and ROS generation capabilities in vitro,the anticancer effects of T@D-siRRS1 NPs on colon cancer were assessed in cell experiments and tumor-bearing mouse models.Results:Both database analysis and clinical specimen testing indicated that RRS1 expression in colon cancer tissues was higher than that in adjacent normal tissues,and high expression was associated with poor prognosis(HR=1.88,95%CI=1.27-2.79,P=0.002).Univariate and multivariate analyses showed that RRS1 mRNA expression was an independent risk factor for overall survival in colon cancer patients(HR=1.354,95%CI=1.447-2.215,P=0.031).Knockdown of RRS1 significantly reduced the proliferation capacity of colon cancer cells and increased apoptosis(both P<0.05).Characterization results showed that the particles were uniform and stable;T@D-siRRS1 NPs entered cells through endocytosis,where TCPP could generate singlet oxygen(1O_2)under ultrasound irradiation.Su

关 键 词：结肠肿瘤 声动力疗法 基因治疗 核糖体生物发生调节蛋白同源物 

分 类 号：R735.3[医药卫生—肿瘤]