2026· American journal of translational research· Vol 18 7, pp.
6069-6082
· 0 citations
Medicine
TL;DR
OtUD6B-AS1 suppresses EAC progression by sponging miR-145-5p and upregulating AIF to induce parthanatos activation, and Targeting this ceRNA regulatory axis may be a therapeutic strategy for EAC.
Abstract
Objectives
To investigate the functional role of OTU deubiquitinase 6B-antisense transcript 1 (OTUD6B-AS1) and its potential association with parthanatos - a poly (ADP-ribose) polymerase 1 (PARP-1)/apoptosis-inducing factor (AIF)-mediated regulated cell death pathway - in esophageal adenocarcinoma (EAC), which remains poorly understood.
Methods
The expression patterns of OTUD6B-AS1, miR-145-5p, and its downstream target AIF protein were investigated in EAC cell lines. Gain- and loss-of-function assays were performed in vitro and in vivo to evaluate biological effects. The competing endogenous RNA (ceRNA) regulatory mechanism was verified using dual-luciferase reporter assays and RNA immunoprecipitation (RIP) assays. Activation of parthanatos was assessed by detecting AIF nuclear translocation and DNA damage levels.
RESULT
OTUD6B-AS1 was reduced significantly, and predominantly localized in the cytoplasm of EAC cells. OTUD6B-AS1 overexpression significantly inhibited EAC cell proliferation, migration, and invasion, and induced cellular apoptosis. Mechanistically, OTUD6B-AS1 functioned as a ceRNA, competitively binging to miR-145-5p and thereby relieving its translational suppression of AIF. This biological pathway promoted parthanatos activation, as evidenced by PARP-1 upregulation, extensive DNA damage (with γ-H2AX deposition), and nuclear translocation of AIF. OTUD6B-AS1-induced antitumor activities were abolished by miR-145-5p overexpression or AIF knockdown. Consistently, OTUD6B-AS1 overexpression substantially suppressed xenograft tumor growth in vivo.
Conclusion
OTUD6B-AS1 suppresses EAC progression by sponging miR-145-5p and upregulating AIF to induce parthanatos activation. Targeting this ceRNA regulatory axis may be a therapeutic strategy for EAC.
Summary Lung adenocarcinoma (LUAD) remains a leading cause of cancer-related death, underscoring the need for an improved molecular understanding. This study investigated the regulatory mechanism of the long non-coding RNA deoxyguanosine kinase antisense RNA 1 (DGUOK-AS1) in LUAD. DGUOK-AS1 was significantly upregulated in LUAD cells and serum samples, and its elevated expression showed a preliminary association with LUAD. Functional experiments demonstrated that DGUOK-AS1 promoted LUAD proliferation and migration both in vitro and in vivo, partly by acting as a competing endogenous RNA for miR-2467-5p to modulate PRMT5 expression. Mechanistically, RNA-binding motif protein 15 (RBM15) enhanced DGUOK-AS1 stability through m6A modification, which in turn enabled heterogeneous nuclear ribonucleoprotein H1 (HNRNPH1) binding in an m6A-dependent manner via its RNA recognition motif 3 (RRM3) domain, promoting degradation. RBM15 knockdown attenuated the malignant phenotype through the miR-2467-5p/PRMT5 axis. These findings reveal an m6A-dependent mechanism governing DGUOK-AS1 stability and provide insights into its contribution to LUAD progression.
Menghao Yang, Jiaen Wu, Youjie Li et al.· iScience· 0 citations
BACKGROUND
Nucleolar protein 2 (NOP2), an RNA methyltransferase that mediates m5C modification, has been implicated in cancer progression; however, its specific role in lung adenocarcinoma (LUAD) remains unclear. This study investigated the functional significance of NOP2 and its regulation of FK506-binding protein 4 (FKBP4) in LUAD pathogenesis.
MATERIALS AND METHODS
The human normal alveolar epithelial cell line HPAEpiC and the LUAD cell lines A549 and H1975 were utilized in this study. In vivo experiments were conducted using male Sprague-Dawley rats. The functional roles of NOP2 and FKBP4 were explored by transfecting LUAD cells with overexpression vectors or small interfering RNAs (siRNAs). Quantitative real-time PCR, western blotting, immunofluorescence, and immunohistochemical staining analyses were used to evaluate NOP2 and FKBP4 expression in cells and tissues. Cell viability and proliferation were assessed using Cell Counting Kit-8 assays and 5-ethynyl-2'-deoxyuridine (EdU) staining. Apoptosis was measured by flow cytometry and TUNEL staining. Transwell assays were conducted to determine cell migration and invasion. Co-immunoprecipitation and RNA pulldown assays were conducted to characterize the interaction between NOP2 and FKBP4.
RESULTS
Analysis of data from The Cancer Genome Atlas (TCGA) database revealed that elevated expression of NOP2 and FKBP4 correlated with poor prognosis in patients with LUAD. Both NOP2 and FKBP4 were significantly upregulated in LUAD cell lines compared to normal controls. Knockdown of NOP2 could inhibit LUAD cell proliferation, migration, and invasion, and enhance apoptosis. In vivo experiments confirmed that NOP2 depletion inhibited tumor growth and enhanced apoptosis. Mechanistically, NOP2 could directly bind to FKBP4 and regulate its expression. Importantly, FKBP4 knockdown reversed the oncogenic effects of NOP2 expression, establishing FKBP4 as a critical downstream effector of NOP2 in LUAD.
CONCLUSION
Our study demonstrates that NOP2 promotes LUAD progression by regulating FKBP4 expression, potentially through m5C methylation, thereby highlighting the NOP2/FKBP4 axis as a potential therapeutic target. Overall, these findings provide new insights into the epigenetic mechanisms driving LUAD aggressiveness.
The ZNF582-AS1/miR-21-5p/miR-516a-3p/CREBRF regulatory axis provides mechanistic insights into EC tumorigenesis and emphasizes the tumor suppressive effect of ZNF582-AS1.
Chen Chen, Fangyuan Chang, Yi Guo et al.· Combinatorial chemistry & hi...· 0 citations
It is indicated that SLC12A9-AS1 is abnormally overexpressed in CRC cells and that its knockdown attenuates malignant phenotypes, at least in part through regulation of miR-139-3p.
Shiyu Zhao, Shuai Zhang, Qiongfeng Tan et al.· Journal of Visualized Experi...· 0 citations
To investigate the effects of the long non-coding RNA (lncRNA) KIAA0125 on the proliferation and apoptosis of the human acute myeloid leukemia subtype M2 (M2-AML) cell line Kasumi-1. Recombinant lentivirus technology and CRISPR/Cas9-SAM technology were used to construct Kasumi-1 cells with downregulated or overexpressed lncKIAA0125, respectively. The cell proliferation rate of each group was determined using the cell counting kit-8 (CCK-8) assay, and the apoptosis rate was determined via flow cytometry. The binding between miR-4640-5p and lncKIAA0125, and between miR-4640-5p and KCNQ1, was subsequently verified using RIP and dual-luciferase assays. Western blotting was used to detect proteins related to the Wnt/β-catenin signaling pathway in each group, and rescue experiments were performed to confirm the role of the lncKIAA0125/miR-4640-5p/KCNQ1 axis in Kasumi-1 cells. Finally, a subcutaneous xenograft tumor model was established to verify the effect of lncKIAA0125 on the growth of Kasumi-1 cells in vivo. Downregulation of lncKIAA0125 inhibited Kasumi-1 cell proliferation and promoted apoptosis, whereas upregulation of lncKIAA0125 had the opposite effect on cell proliferation. RIP and dual-luciferase assays confirmed that lncKIAA0125 directly bound to miR-4640-5p and that miR-4640-5p also bound to KCNQ1. Moreover, downregulation of lncKIAA0125 inhibited KCNQ1 expression at both the mRNA and protein levels and reduced signaling related to the Wnt/β-catenin pathway. Conversely, upregulation of lncKIAA0125 produced the opposite effects. Finally, in vivo findings demonstrated that downregulating lncKIAA0125 expression suppressed the proliferation of Kasumi-1 cells in nude mice. lncKIAA0125 is highly expressed in AML cells and appears to promote the proliferation of Kasumi-1 cells via the miR-4640-5p/KCNQ1 axis and was associated with changes in the Wnt/β-catenin signaling pathway.
Huali Hu, Yuancheng Liu, Fahua Deng et al.· Scientific Reports· 0 citations
Although Esketamine, an N-methyl-D-aspartate (NMDA) receptor antagonist, is widely used for anesthesia and analgesia and has shown anticancer activity, its effects on lung adenocarcinoma remain unclear. Circular RNAs have been implicated in the development and progression of lung adenocarcinoma. Therefore, this study aims to elucidate whether esketamine can influence lung adenocarcinoma progression by interfering with the circular RNA hsa_circ_0022392.
Expression levels of hsa_circ_0022392, miR-548b-3p, and microtubule-associated protein 7 (MAP7), as well as transfection efficiency, were assessed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis. Cell viability was assessed using the Cell Counting Kit-8 assay. 5-ethynyl-2’-deoxyuridine (EdU) assay was employed to examine cell proliferation. Transwell assays were employed for analyzing cell migration and invasion capabilities. Intracellular Fe²⁺ levels, malondialdehyde (MDA), glutathione (GSH), and reactive oxygen species (ROS) were measured using commercial kits. Lactate dehydrogenase (LDH) release assays were adopted to evaluate CD8⁺ T-cell cytotoxicity. Cytokine levels were measured via ELISA. Bioinformatics analysis combined with RNA immunoprecipitation and dual-luciferase reporter assays were employed to confirm the target relationship between miR-548b-3p and hsa_circ_0022392 or MAP7.
Esketamine significantly inhibited proliferation, migration, and invasion (all
P
< 0.05), induced ferroptosis (
P
< 0.05), and suppressed immune evasion (
P
< 0.05). Notably, hsa_circ_0022392 expression was upregulated in lung adenocarcinoma tissues and cell lines (
P
< 0.01), and esketamine downregulated its levels (
P
< 0.05). Overexpressed hsa_circ_0022392 notably reversed esketamine’s inhibitory effect on lung adenocarcinoma cell progression (
P
< 0.05). Further studies revealed that hsa_circ_0022392 targeted and bound to miR-548b-3p, thereby upregulating MAP7 expression (
P
< 0.05). Interfering with this signaling axis effectively weakened the inhibitory effect of esketamine on lung adenocarcinoma progression (
P
< 0.05).
In summary, esketamine promotes ferroptosis in lung adenocarcinoma cells through regulating the hsa_circ_0022392/miR-548b-3p/MAP7 axis and inhibits immune evasion, providing novel molecular targets and therapeutic strategies for the diagnosis and treatment of lung adenocarcinoma.
Fei Chai, Yang Zhang, Xing-Jia Chen et al.· Hereditas· 0 citations
