可编辑DNA纳米花抑制LLC细胞增殖、迁移及侵袭的实验研究  

An experimental study of editable DNA nanoflowers inhibiting the proliferation,migration and invasion of Lewis lung carcinoma cells

作　　者：曹宇超 廖洪建 胡璨 郭佳俊 杜永洪[1] Cao Yuchao;Liao Hongjian;Hu Can;Guo Jiajun;Du Yonghong(State Key Laboratory of Ultrasound in Medicine and Engineering,College of Biomedical Engineering,Chongqing Medical University)

机构地区：[1]重庆医科大学生物医学工程学院/超声医学工程国家重点实验室,重庆400016

出　　处：《重庆医科大学学报》2024年第9期1105-1112,共8页Journal of Chongqing Medical University

基　　金：重庆市技术创新与应用发展专项面上资助项目(编号:cstc2019jscx-msxmX0255)。

摘　　要：目的:探究载有阿霉素(doxorubicin,DOX)和缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)反义寡核苷酸的DNA纳米花(DNA nanoflower,DF)对小鼠肺癌(lewis lung carcinoma,LLC)细胞增殖、迁移及侵袭能力的影响。方法:通过滚环扩增反应(rolling circle amplification,RCA)合成含有HIF-1α反义寡核苷酸的DHA-DF,并检测其物理特性;负载DOX后获得载DOX纳米花(DOX-loaded DNA nanoflower,DDF)。通过细胞计数试剂(cell counting kit-8,CCK-8)评价DHA-DDF对LLC细胞活性的影响;通过划痕实验和Transwell小室实验检测DHA-DDF对LLC细胞迁移及侵袭能力的影响;通过蛋白质印记实验和定量聚合酶链反应检测LLC细胞HIF-1α的表达水平。结果:DHA-DF为多孔花瓣状球形颗粒,粒径为(421.26±7.90)nm;与游离DOX和DA-DDF相比,经DHA-DDF处理后的LLC细胞HIF-1α蛋白及mRNA的表达量明显下降(均P<0.05),且其细胞活性、划痕愈合面积及侵袭细胞数目明显减少(均P<0.05)。结论:DHA-DDF能有效降低HIF-1α的表达并抑制LLC细胞的增殖、迁移和侵袭,有望为临床肿瘤治疗提供新的思路。Objective:To investigate the effects on the proliferation,migration and invasion of Lewis lung carcinoma(LLC)cells in⁃duced by DNA nanoflowers(DFs)carrying doxorubicin(DOX)and hypoxia inducible factor-1α(HIF-1α)antisense oligonucleotides.Methods:DFs containing HIF-1αantisense oligonucleotides(DHA-DF)were synthesized by rolling circle amplification,and the physical properties of DHA-DF were examined.DOX was loaded into DFs to form DDF.We evaluated the cell viability of LLC cells treated with DHA-DDF by using Cell Counting Kit-8;assessed the effects of DHA-DDF on the migration and invasion of LLC cells by using scratch assay and Transwell assay;and measured the expression level of HIF-1αin LLC cells by Western blot and quantitative polymerase chain reaction.Results:DHA-DF appeared flower-like porous spherical particles with a size of(421.26±7.90)nm.Com⁃pare with free DOX and DA-DDF,DHA-DDF significantly decreased the mRNA and protein expression of HIF-1αin LLC cells,and significantly reduced cell viability,the area of wound healed and the number of invasive cells(all P<0.05).Conclusion:DHA-DDF could effectively reduce the expression of HIF-1α,and inhibit the proliferation,migration and invasion of LLC cells,providing new ideas for clinical treatment of tumors.

关 键 词：DNA纳米花 缺氧 肺癌 反义寡核苷酸 

分 类 号：R734.2[医药卫生—肿瘤]