金纳米颗粒通过调节三阴性乳腺癌细胞中的 microRNA-155-5p，减弱干扰素-γ诱导的 SOCS1 表达和 NF-κB p65/50 活性的激活。

Gold nanoparticles attenuate the interferon-γ induced SOCS1 expression and activation of NF-κB p65/50 activity via modulation of microRNA-155-5p in triple-negative breast cancer cells.

机构信息

Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka, Aljouf, Saudi Arabia.

Department of Medical Biochemistry, College of Medicine, Qassim University, Buraidah, Saudi Arabia.

出版信息

Front Immunol. 2023 Aug 31;14:1228458. doi: 10.3389/fimmu.2023.1228458. eCollection 2023.

DOI:10.3389/fimmu.2023.1228458

PMID:37720228

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10500308/

Abstract

OBJECTIVE

Triple-negative breast cancer (TNBC) is a very aggressive form of cancer that grows and spreads very fast and generally relapses. Therapeutic options of TNBC are limited and still need to be explored completely. Gold nanoparticles conjugated with citrate (citrate-AuNPs) are reported to have anticancer potential; however, their role in regulating microRNAs (miRNAs) in TNBC has never been investigated. This study investigated the potential of citrate-AuNPs against tumorigenic inflammation via modulation of miRNAs in TNBC cells.

METHODS

Gold nanoparticles were chemically synthesized using the trisodium-citrate method and were characterized by UV-Vis spectrophotometry and dynamic light scattering studies. Targetscan bioinformatics was used to analyze miRNA target genes. Levels of miRNA and mRNA were quantified using TaqMan assays. The pairing of miRNA in 3'untranslated region (3'UTR) of mRNA was validated by luciferase reporter clone, containing the entire 3'UTR of mRNA, and findings were further re-validated via transfection with miRNA inhibitors.

RESULTS

Newly synthesized citrate-AuNPs were highly stable, with a mean size was 28.3 nm. The data determined that hsa-miR155-5p is a direct regulator of SOCS1 (suppressor-of-cytokine-signaling) expression and citrate-AuNPs inhibits SOCS1 mRNA/protein expression via modulating hsa-miR155-5p expression. Transfection of TNBC MDA-MB-231 cells with anti-miR155-5p markedly increased SOCS1 expression (p<0.001), while citrate-AuNPs treatment significantly inhibited anti-miR155-5p transfection-induced SOCS1 expression (p<0.05). These findings were validated by IFN-γ-stimulated MDA-MB-231 cells. Moreover, the data also determined that citrate-AuNPs also inhibit IFN-γ-induced NF-κB p65/p50 activation in MDA-MB-231 cells transfected with anti-hsa-miR155-5p.

CONCLUSION

Newly generated citrate-AuNPs were stable and non-toxic to TNBC cells. Citrate-AuNPs inhibit IFN-γ-induced SOCS1 mRNA/protein expression and deactivate NF-κB p65/50 activity via negative regulation of hsa-miR155-5p. These novel pharmacological actions of citrate-AuNPs on IFN-γ-stimulated TNBC cells provide insights that AuNPs inhibit IFN-γ induced inflammation in TNBC cells by modulating the expression of microRNAs.

摘要

目的

三阴性乳腺癌（TNBC）是一种侵袭性很强的癌症，生长和扩散非常迅速，通常会复发。TNBC 的治疗选择有限，仍需进一步探索。已报道金纳米粒子与柠檬酸盐结合（柠檬酸盐-AuNPs）具有抗癌潜力；然而，其在调节 TNBC 细胞中的 microRNAs（miRNAs）方面的作用从未被研究过。本研究通过调节 TNBC 细胞中的 miRNAs 来研究柠檬酸盐-AuNPs 对肿瘤发生性炎症的潜在作用。

方法

使用柠檬酸三钠法化学合成金纳米粒子，并通过紫外-可见分光光度法和动态光散射研究进行表征。使用 Targetscan 生物信息学分析 miRNA 靶基因。使用 TaqMan 测定法定量测定 miRNA 和 mRNA 的水平。通过包含 mRNA 全长 3'UTR 的荧光素酶报告克隆验证 miRNA 在 mRNA 3'UTR 中的配对，并通过 miRNA 抑制剂转染进一步验证。

结果

新合成的柠檬酸盐-AuNPs 非常稳定，平均粒径为 28.3nm。数据表明 hsa-miR155-5p 是 SOCS1（细胞因子信号转导抑制因子）表达的直接调节剂，柠檬酸盐-AuNPs 通过调节 hsa-miR155-5p 的表达抑制 SOCS1 mRNA/蛋白的表达。用抗 hsa-miR155-5p 转染 TNBC MDA-MB-231 细胞可显著增加 SOCS1 表达（p<0.001），而柠檬酸盐-AuNPs 处理可显著抑制抗 hsa-miR155-5p 转染诱导的 SOCS1 表达（p<0.05）。这一发现通过 IFN-γ 刺激的 MDA-MB-231 细胞得到了验证。此外，数据还表明，柠檬酸盐-AuNPs 还抑制转染抗 hsa-miR155-5p 的 MDA-MB-231 细胞中 IFN-γ 诱导的 NF-κB p65/p50 激活。

结论

新生成的柠檬酸盐-AuNPs 稳定且对 TNBC 细胞无毒。柠檬酸盐-AuNPs 通过负调控 hsa-miR155-5p 抑制 IFN-γ 诱导的 SOCS1 mRNA/蛋白表达并使 NF-κB p65/50 失活。柠檬酸盐-AuNPs 对 IFN-γ 刺激的 TNBC 细胞的这些新的药理作用表明，AuNPs 通过调节 microRNAs 的表达抑制 IFN-γ 诱导的 TNBC 细胞炎症。