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壳聚糖纳米颗粒介导的抗微小RNA-324-5p通过调控GLI1表达对降低卵巢癌细胞增殖的作用。

Chitosan nanoparticle-mediated effect of antimiRNA-324-5p on decreasing the ovarian cancer cell proliferation by regulation of GLI1 expression.

作者信息

Ysrafil Ysrafil, Astuti Indwiani

机构信息

Department of Pharmacy, Politeknik Kesehatan Kementerian Kesehatan Gorontalo, Gorontalo, Indonesia.

Department of Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.

出版信息

Bioimpacts. 2022;12(3):195-202. doi: 10.34172/bi.2021.22119. Epub 2021 Nov 29.

Abstract

MicroRNAs (miRNAs) are short-sequence RNAs that regulate gene expression by targeting messenger RNAs (mRNAs). Recent studies reveal that miRNA-324-5p plays an important role in worsening the ovarian cancer prognosis when the expression is very high. This study aimed to develop a miRNA targeted therapy by targeting the miRNA-324-5p function as a miRNA-324-5p inhibitor. Chitosan nanoparticles were used for antimiRNA-324-5p delivery into SKOV3 cell lines formulated by ionic gelation method. Antiproliferative effect of CS-NPs-antimiRNA was assessed by the MTT Assay. A mechanism study assessed the anticancer effect of the formula. analysis used miRTar.Human and StarmiRDB combined with Genecard to predict the target genes of antimiR. Hawkdock web server was used to analyze protein-protein interactions that were further validated by quantitative polymerase chain reaction (qPCR). The results of qPCR analysis showed endogenous miRNA-324-5p decreased after 24-hour transfection of antagonist miRNA. Furthermore, the MTT assay results showed that antimiRNA was able to inhibit SKOV3 cell proliferation (80 nM 68.13%, < 0.05). analysis found miRNA-324-5p can regulate MEN1 and indirectly repress Gli1 mRNA. Validation results confirmed antimiR can decrease GLI1 mRNA expression. Our results showed antimiRNA-324-5p can act as a microRNA-based therapy to inhibit ovarian cancer proliferation by the reduction of GLI1 expression.

摘要

微小RNA(miRNA)是一类短序列RNA,通过靶向信使RNA(mRNA)来调节基因表达。最近的研究表明,当miRNA-324-5p表达非常高时,它在恶化卵巢癌预后方面发挥重要作用。本研究旨在通过靶向miRNA-324-5p的功能,开发一种作为miRNA-324-5p抑制剂的miRNA靶向治疗方法。壳聚糖纳米颗粒通过离子凝胶法制备,用于将抗miRNA-324-5p递送至SKOV3细胞系。通过MTT法评估CS-NPs-抗miRNA的抗增殖作用。机制研究评估了该配方的抗癌效果。分析使用miRTar.Human和StarmiRDB并结合Genecard来预测抗miR的靶基因。使用Hawkdock网络服务器分析蛋白质-蛋白质相互作用,并通过定量聚合酶链反应(qPCR)进一步验证。qPCR分析结果显示,在转染拮抗剂miRNA 24小时后,内源性miRNA-324-5p减少。此外,MTT试验结果表明,抗miRNA能够抑制SKOV3细胞增殖(80 nM时为68.13%,<0.05)。分析发现miRNA-324-5p可以调节MEN1并间接抑制Gli1 mRNA。验证结果证实抗miR可以降低GLI1 mRNA表达。我们的结果表明,抗miRNA-324-5p可以作为一种基于微小RNA的治疗方法,通过降低GLI1表达来抑制卵巢癌增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b3a/9124874/0a1b8129065b/bi-12-195-g001.jpg

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