聚乳酸-羟基乙酸共聚物负载siRNA-circCALN1治疗鼻咽癌的初步研究  

Preliminary study on the treatment of nasopharyngeal carcinoma with poly lactic-co-glycolic acid loaded siRNA-circCALN1

作　　者：江浩 张鑫 舒能何 陈舒华 曲延玉 龚媛 JIANG Hao;ZHANG Xin;SHU Neng-he;CHEN Shu-hua;QU Yan-yu;GONG Yuan(Department of Pathology,The Second People's Hospital of Foshan/Affiliated Foshan Hospital of Southern Medical University,Foshan,Guangdong 528000;Shuidong Health Center of Zhanggong District,Ganzhou,Jiangxi 341000;Department of Otolaryngology,The Second People's Hospital of Foshan/Affiliated Foshan Hospital of Southern Medical University;Department of General Practice,The Second People's Hospital of Foshan/Affiliated Foshan Hospital of Southern Medical University,Foshan,Guangdong 528000)

机构地区：[1]南方医科大学附属佛山医院/佛山市第二人民医院病理科,广东佛山528000 [2]赣州市章贡区水东镇卫生院,江西赣州341000 [3]南方医科大学附属佛山医院/佛山市第二人民医院耳鼻咽喉科 [4]南方医科大学附属佛山医院/佛山市第二人民医院全科,广东佛山528000

出　　处：《赣南医学院学报》2023年第4期333-338,共6页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：2020年佛山市自筹经费类科技计划项目(医学类科技攻关项目)(2020001005658)。

摘　　要：目的:为了提高疗效,我们在前期研究的基础上开发了一种小干扰核糖核酸(Small interfering RNA,siRNA)和聚乳酸-羟基乙酸共聚物[poly(lactic-co-glycolic acid),PLGA]结合点的纳米复合药物,为开发新的抗鼻咽癌(Nasopharyngeal carcinoma,NPC)药物提供新方法和思路。方法:设计并合成三个针对circRNA CALN1后剪接区的siRNA序列(siRNA1 circ CALN1、siRNA2 circ CALN1和siRNA3 circ CALN1)和非功能性siRNA(Nonfunctional siRNA,siRNA NC),通过RT-PCR进行抑制circ CALN1探究。对合成好的PLGA-siRNA circ CALN1进行SEM、TEM、DLS、Zeta电位等表征。通过绘制标准曲线,计算siRNA负载率。通过CCK-8对两种鼻咽癌抗癌作用进行验证。结果:通过RT-PCR验证了siRNA3 circ CALN1对circ CALN1抑制作用最强。通过SEM和TEM进行形貌表征,PLGA-siRNA-circ CALN1呈均匀球形。DLS显示水合粒径205 nm左右,Zeta电位-14 mV,说明已经成功负载。通过绘制标准曲线,计算siRNA负载率为(92.3±5.4)%。在培养24、48、72、96 h后,与PLGA-siRNA NC组和对照组比较,PLGA-siRNA3 circ CALN1可显著抑制鼻咽癌细胞(CNE1和HNE1)的增殖。结论:本研究开发的治疗鼻咽癌的抗癌药物获得初步成功,开发了鼻咽癌的新靶点,丰富了抗癌药物的工具箱。Objective:In order to improve the efficacy of the treatment of nasopharyngeal carcinoma,a small interfering RNA(siRNA)and poly(lactic-co-glycolic acid)was developed on the basis of our previous study,PLGA combined nanocomposite drugs to provide new methods and ideas for the development of new anti-nasopharyngeal carcinoma drugs.Methods:We designed and synthesized three siRNAs targeting the posterior splice region of circ CALN1(siRNA1 circ CALN1,siRNA2 circ CALN1 and siRNA3-circ CALN1)and nonfunctional siRNAs(siRNA NC)Inhibition of circ CALN1 was investigated by RT-PCR.The synthesized PLGA-siRNA-circ CALN1 was characterized by SEM,TEM,DLS and Zeta potential.The siRNA loading rate was calculated by drawing standard curves.The anticancer effect of two nasopharyngeal carcinomas was verified by CCK-8.Results:siRNA3-circ CALN1 showed the strongest inhibitory effect on circ CALN1 by RT-PCR.The morphology of PLGA-siRNA-circ CALN1 was characterized by SEM and TEM.DLS shows that the hydrated particle size is about 205 nm and the Zeta potential is-14 mV,indicating that the load has been successfully loaded.The siRNA loading rate was calculated as(92.3±5.4)%by drawing the standard curve.After 24 hours,48 hours,72 hours and 96 hours of culture,PLGA-siRNA3 circ CALN1 significantly inhibited the proliferation of nasopharyngeal carcinoma cells(CNE1 and HNE1)compared with PLGA-siRNA NC group and control group.Conclusion:The anticancer drugs developed in this study have achieved initial success in the treatment of nasopharyngeal carcinoma,which has developed new targets for nasopharyngeal carcinoma and enriched the toolbox of anticancer drugs.

关 键 词：鼻咽癌 环状核糖核酸CALN1 小干扰核糖核酸 聚乳酸-羟基乙酸共聚物 纳米粒子 

分 类 号：R766.3[医药卫生—耳鼻咽喉科]