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 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.