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

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 seq...

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Main Authors: Omid Nikkhah, Behzad Einollahi, Mosa Asadi, Mohammad Heiat, Kiavash Hushmandi
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Biochemistry and Biophysics Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2405580825001281
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author Omid Nikkhah
Behzad Einollahi
Mosa Asadi
Mohammad Heiat
Kiavash Hushmandi
author_facet Omid Nikkhah
Behzad Einollahi
Mosa Asadi
Mohammad Heiat
Kiavash Hushmandi
author_sort Omid Nikkhah
collection DOAJ
description 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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spelling doaj-art-c538db355b774c2d9d10ec6d4bdc8d4a2025-08-20T02:17:09ZengElsevierBiochemistry and Biophysics Reports2405-58082025-06-014210204110.1016/j.bbrep.2025.102041Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model studyOmid Nikkhah0Behzad Einollahi1Mosa Asadi2Mohammad Heiat3Kiavash Hushmandi4Nephrology 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, IranNephrology and Urology Research Center, Clinical Sciences Institute, Baqiyatallah University of Medical Sciences, Tehran, IranNephrology and Urology Research Center, Clinical Sciences Institute, Baqiyatallah University of Medical Sciences, Tehran, IranBaqiyatallah Research Center for Gastroenterology and Liver Diseases (BRCGL), Clinical Sciences Institute, Baqiyatallah University of Medical Sciences, Tehran, IranNephrology and Urology Research Center, Clinical Sciences Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran; Corresponding author.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.http://www.sciencedirect.com/science/article/pii/S2405580825001281AntisenseBladder cancermiR-21miR-373Nanoliposome
spellingShingle Omid Nikkhah
Behzad Einollahi
Mosa Asadi
Mohammad Heiat
Kiavash Hushmandi
Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model study
Biochemistry and Biophysics Reports
Antisense
Bladder cancer
miR-21
miR-373
Nanoliposome
title Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model study
title_full Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model study
title_fullStr Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model study
title_full_unstemmed Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model study
title_short Gene network changes after exposure to nanoliposomes containing antisense miR-21 and miR-373 in bladder cancer Cells: An in vitro model study
title_sort gene network changes after exposure to nanoliposomes containing antisense mir 21 and mir 373 in bladder cancer cells an in vitro model study
topic Antisense
Bladder cancer
miR-21
miR-373
Nanoliposome
url http://www.sciencedirect.com/science/article/pii/S2405580825001281
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