Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model study

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2025-05-12 DOI:10.1016/j.bbrep.2025.102041

Omid Nikkhah , Behzad Einollahi , Mosa Asadi , Mohammad Heiat , Kiavash Hushmandi

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引用次数: 0

Abstract

Aims

This study aimed to examine the changes in gene expression profiles of the bladder cancer cell line (HTB-9) after exposure with nanoliposomes (NLs) containing antisense miR-21, antisense miR-373, or a combination of both antisense miR-21 and antisense miR-373 oligonucleotides.

Methods

The sequence of miR-21 and miR-373 was obtained from the NCBI, and the optimal corresponding antisense oligonucleotides (ASOs) were selected and synthesized using the Oligowalk online server. After encapsulating the ASOs in liposomes and characterizing them, the liposomal ASOs were incubated with the target cells for 24 h at 37 °C. Following incubation, total RNA was extracted, and cDNA was synthesized. The expression levels of miR-21, miR-373, and eight additional core genes (STK38L: Serine/threonine-protein kinase 38-like; PCDH19: Protocadherin-19; YOD1: Ubiquitin thioesterase OTU1; PRDM11: PR domain-containing protein 11; CROT: Peroxisomal carnitine O-octanoyltransferase; LATS2: Serine/threonine-protein kinase; ZNF845: Zinc finger protein 845; ZC3H6: Zinc finger CCCH domain-containing protein 6) were then analyzed using quantitative Reverse Transcriptase - PCR (qRT-PCR).

Results

ASOmiR-21 (AUCUCAUGGCAACACCAGU) and ASOmiR-373 (AAGUGCUUCGAUUUUGGGG) nucleotides were used in this study, respectively. Data analysis revealed that the expression levels of miR-21 and miR-373 were significantly reduced in HTB-9 cells exposed to nanoliposomal ASOs (NL-ASOs) with sizes ranging from 100 ± 5 to 260 ± 10 nm, compared to the control groups. Furthermore, HTB-9 cells exposed simultaneously to both liposomal ASOs (NL-ASOmiR-21+ASOmiR-373) exhibited a greater reduction in miR-21 and miR-373 expression. Additionally, all studied genes (STK38L, PCDH19, YOD1, PRDM11, CROT, LATS2, ZNF845, ZC3H6) showed significant decreases in expression in HTB-9 cells exposed to NL-ASOs across all experimental designs.

Conclusions

The results demonstrated that miR-21 and miR-373 play crucial roles in gene expression and that their inhibition can significantly impact the expression profile of a gene network in bladder cancer. Therefore, to regulate the expression of a gene network in bladder cancer, we can use antimir technology as an effective strategy.

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膀胱癌细胞暴露于含有反义miR-21和miR-373的纳米脂质体后基因网络的变化：一项体外模型研究

目的本研究旨在检测膀胱癌细胞系（HTB-9）暴露于含有反义miR-21、反义miR-373或反义miR-21和反义miR-373寡核苷酸的纳米脂质体（NLs）后基因表达谱的变化。方法从NCBI中获取miR-21和miR-373的序列，通过Oligowalk在线服务器选择相应的最优反义寡核苷酸（ASOs）进行合成。将ASOs包封在脂质体中并进行表征后，将脂质体ASOs与靶细胞在37℃下孵育24 h。孵育后提取总RNA，合成cDNA。miR-21、miR-373和另外8个核心基因(STK38L：丝氨酸/苏氨酸蛋白激酶38样；PCDH19: Protocadherin-19;YOD1：泛素硫酯酶OTU1；PRDM11: PR结构域含蛋白11；CROT：过氧化物酶体肉碱o -辛烷基转移酶；LATS2：丝氨酸/苏氨酸蛋白激酶；ZNF845：锌指蛋白845；ZC3H6：锌指CCCH结构域蛋白6)采用定量逆转录酶-PCR （qRT-PCR）分析。结果本研究分别使用了asomir -21 （AUCUCAUGGCAACACCAGU）和ASOmiR-373 （AAGUGCUUCGAUUUUGGGG）核苷酸。数据分析显示，与对照组相比，暴露于大小为100±5至260±10 nm的纳米脂质体ASOs （NL-ASOs）的HTB-9细胞中miR-21和miR-373的表达水平显著降低。此外，同时暴露于两种脂质体ASOs （NL-ASOmiR-21+ASOmiR-373）的HTB-9细胞显示出miR-21和miR-373表达的更大降低。此外，所有研究基因（STK38L、PCDH19、YOD1、PRDM11、CROT、LATS2、ZNF845、ZC3H6）在所有实验设计中暴露于NL-ASOs的HTB-9细胞中的表达均显著降低。结论miR-21和miR-373在膀胱癌基因表达中起着至关重要的作用，抑制miR-21和miR-373可显著影响膀胱癌基因网络的表达谱。因此，调控膀胱癌中某一基因网络的表达，我们可以将抗mir技术作为一种有效的策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.