Rossetto Isabela Maria Urra, Alves Letícia F, Marson Leonardo A, Geraldo Murilo V, Santos Felipe R, Montico Fábio, Cagnon Valéria H A
Department of Structural and Functional Biology, University of Campinas (UNICAMP), Campinas, SP, Brazil.
Department of Structural and Functional Biology, University of Campinas, 255 Monteiro Lobato St, Campinas, SP, 13083-862, Brazil.
J Mol Histol. 2025 Jan 13;56(1):69. doi: 10.1007/s10735-024-10341-y.
This study investigated tempol action on genes and miRNAs related to NFκB pathway in androgen dependent or independent cell lines and in TRAMP model in the early and late-stages of cancer progression. A bioinformatic search was conducted to select the miRNAs to be measured based on the genes of interest from NFκB pathway. The miR-let-7c-5p, miR-26a-5p and miR-155-5p and five target genes (BCL2, BCL2L1, RELA, TNF, PTGS2) were chosen for RT-PCR and gene enrichment analyses. In vitro, PC-3 and LNCaP cells were exposed, respectively, to 1.0 or 2.0 mM of tempol during 48 h. In vivo, five experimental groups were evaluated regarding tempol effects in the early (CT12 and TPL12 groups) and late-stages (CT20, TPL20-I and TLP20-II) of PCa development. TPL groups were treated with 50 mg/kg or 100 mg/kg of tempol. The ventral lobe of the prostate and the plasma was collected. Tempol treatment increased miRs expression in PC-3 and LNCaP. For both cell lines, tempol decreased RELA expression. In TRAMP model, tempol increased miRNA expression in prostate for all treated groups. Tempol upregulated the miRNA expressions related to the NFκB pathway in the prostate tissue and human tumor cell lines. Their increase is mainly linked to increased cell death and delayed CaP aggressivenes. Thus, tempol's capacity for miRNA-mediated gene silencing to decrease tissue proliferation and cell survival processes is part of its tissue mechanics.
本研究调查了Tempol对雄激素依赖或非依赖细胞系以及前列腺癌（TRAMP）模型癌症进展早期和晚期中与NFκB通路相关的基因和微小RNA（miRNA）的作用。基于NFκB通路的目标基因进行了生物信息学搜索，以选择要检测的miRNA。选择了miR-let-7c-5p、miR-26a-5p和miR-155-5p以及五个靶基因（BCL2、BCL2L1、RELA、TNF、PTGS2）进行逆转录聚合酶链反应（RT-PCR）和基因富集分析。在体外，PC-3和LNCaP细胞分别在48小时内暴露于1.0或2.0 mM的Tempol。在体内，评估了五个实验组Tempol在前列腺癌（PCa）发展早期（CT12和TPL12组）和晚期（CT20、TPL20-I和TLP20-II）的作用。TPL组用50 mg/kg或100 mg/kg的Tempol处理。收集前列腺腹叶和血浆。Tempol处理增加了PC-3和LNCaP中miR的表达。对于这两种细胞系，Tempol降低了RELA的表达。在TRAMP模型中，Tempol增加了所有治疗组前列腺中的miRNA表达。Tempol上调了前列腺组织和人肿瘤细胞系中与NFκB通路相关的miRNA表达。它们的增加主要与细胞死亡增加和前列腺癌侵袭延迟有关。因此，Tempol通过miRNA介导的基因沉默来降低组织增殖和细胞存活过程的能力是其组织作用机制的一部分。
