Nikkhah Omid, Einollahi Behzad, Asadi Mosa, Heiat Mohammad, Hushmandi Kiavash
Nephrology and Urology Research Center, Clinical Sciences Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Department of Urology, Clinical Research Development Unit of Shahid Madani Hospital, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.
Biochem Biophys Rep. 2025 May 12;42:102041. doi: 10.1016/j.bbrep.2025.102041. eCollection 2025 Jun.
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.
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 (: Serine/threonine-protein kinase 38-like; : Protocadherin-19; Ubiquitin thioesterase OTU1; PR domain-containing protein 11; Peroxisomal carnitine -octanoyltransferase; Serine/threonine-protein kinase; Zinc finger protein 845; Zinc finger CCCH domain-containing protein 6) were then analyzed using quantitative Reverse Transcriptase - PCR (qRT-PCR).
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 (, , , , , , , ) showed significant decreases in expression in HTB-9 cells exposed to NL-ASOs across all experimental designs.
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.
本研究旨在检测膀胱癌细胞系（HTB - 9）在暴露于含有反义miR - 21、反义miR - 373或反义miR - 21与反义miR - 373寡核苷酸组合的纳米脂质体（NLs）后基因表达谱的变化。
从NCBI获取miR - 21和miR - 373的序列，并使用Oligowalk在线服务器选择并合成最佳对应的反义寡核苷酸（ASOs）。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°C下孵育24小时。孵育后，提取总RNA并合成cDNA。然后使用定量逆转录 - PCR（qRT - PCR）分析miR - 21、miR - 373以及另外八个核心基因（：丝氨酸/苏氨酸蛋白激酶38样；：原钙黏蛋白 - 19；泛素硫酯酶OTU1；含PR结构域蛋白11；过氧化物酶体肉碱 - 辛酰转移酶；丝氨酸/苏氨酸蛋白激酶；锌指蛋白845；含锌指CCCH结构域蛋白6）的表达水平。
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，暴露于尺寸范围为100±5至260±10nm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细胞中，miR - 21和miR - 373的表达降低更为明显。此外，在所有实验设计中，暴露于NL - ASOs的HTB - 9细胞中所有研究的基因（，，，，，，，）的表达均显著下降。
结果表明，miR - 21和miR - 373在基因表达中起关键作用，它们的抑制可显著影响膀胱癌基因网络的表达谱。因此，为调控膀胱癌基因网络的表达，我们可以使用抗miR技术作为一种有效策略。
