可编辑DNA纳米花抑制LLC细胞增殖、迁移及侵袭的实验研究

曹宇超, 廖洪建, 胡璨, 郭佳俊, 杜永洪

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重庆医科大学学报 ›› 2024, Vol. 49 ›› Issue (09) : 1105-1112. DOI: 10.13406/j.cnki.cyxb.003578
基础研究

可编辑DNA纳米花抑制LLC细胞增殖、迁移及侵袭的实验研究

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

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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。

Abstract

Objective To investigate the effects on the proliferation,migration and invasion of Lewis lung carcinoma(LLC) cells induced by DNA nanoflowers(DFs) carrying doxorubicin(DOX) and hypoxia inducible factor-1α(HIF-1α) antisense oligonucleotides. Methods DFs containing HIF-1α antisense 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-1α in 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. Compare with free DOX and DA-DDF,DHA-DDF significantly decreased the mRNA and protein expression of HIF-1α in 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

Key words

DNA nanoflower / hypoxia / lung cancer / antisense oligonucleotide

中图分类号

R734.2

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曹宇超 , 廖洪建 , 胡璨 , . 可编辑DNA纳米花抑制LLC细胞增殖、迁移及侵袭的实验研究. 重庆医科大学学报. 2024, 49(09): 1105-1112 https://doi.org/10.13406/j.cnki.cyxb.003578
Cao Yuchao, Liao Hongjian, Hu Can, et al. An experimental study of editable DNA nanoflowers inhibiting the proliferation,migration and invasion of Lewis lung carcinoma cells[J]. Journal of Chongqing Medical University. 2024, 49(09): 1105-1112 https://doi.org/10.13406/j.cnki.cyxb.003578

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基金

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

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