Journal: Oncology Letters

Article Title: METTL5 reprograms glycolytic metabolism and promotes non-small cell lung cancer progression by modifying PGAM1

doi: 10.3892/ol.2026.15609

Figure Lengend Snippet: Western blot analysis of GLUT1 protein levels under different experimental conditions. (A) GLUT1 protein expression was significantly reduced in PGAM1-knockdown cell lines and significantly elevated in PGAM1-overexpressing cell lines. (B) GLUT1 protein expression was significantly reduced in METTL5-knockdown cell lines and significantly elevated in METTL5-overexpressing cell lines. (C) GLUT1 protein expression was upregulated in METTL5-silenced cells co-transfected with PGAM1 overexpression plasmid. β-actin was used as the loading control. ***P<0.001 and ****P<0.0001. GLUT1, glucose transporter type 1; PGAM1, phosphoglycerate mutase 1; METTL5, methyltransferase 5; NC, negative control; oe, overexpression; si, small interfering RNA.

Article Snippet: METTL5- and PGAM1-overexpression plasmids were purchased from Heyuan Liji (Shanghai) Biotechnology Co., Ltd.

Techniques: Western Blot, Expressing, Knockdown, Transfection, Over Expression, Plasmid Preparation, Control, Negative Control, Small Interfering RNA

Journal: Frontiers in Oncology

Article Title: miR-2110 orchestrates ERK–ELK1 transcriptional repression to induce cell-cycle arrest and enhance cytarabine sensitivity in acute myeloid leukemia

doi: 10.3389/fonc.2026.1833649

Figure Lengend Snippet: Identification of miR-2110 target genes and functional enrichment analysis. (A) Overlap of predicted target genes of miR-2110 from multiple databases. (B) Top-ranked candidate target genes based on integrated scoring. (C–D) GO and KEGG enrichment analyzes of predicted target genes, suggesting potential involvement in transcriptional regulation and signaling pathways. (E) Schematic illustration of predicted binding sites between miR-2110 and ELK1 3′UTR.

Article Snippet: miR-2110 mimics, negative control (NC) oligonucleotides, and ELK1 overexpression plasmids, along with their corresponding control vectors, were purchased from Sangon Biotech (Shanghai, China).

Techniques: Functional Assay, Protein-Protein interactions, Binding Assay

Journal: Frontiers in Oncology

Article Title: miR-2110 orchestrates ERK–ELK1 transcriptional repression to induce cell-cycle arrest and enhance cytarabine sensitivity in acute myeloid leukemia

doi: 10.3389/fonc.2026.1833649

Figure Lengend Snippet: Mapping of functional miR-2110 binding sites within the ELK1 3′UTR. (A) Schematic illustration of mutant ELK1 3′UTR reporter constructs. Four individual mutants (MUT#1–MUT#4) were generated by introducing nucleotide substitutions into each predicted miR-2110 binding region (352–359, 593–599, 1226–1233, and 3985–3991, respectively), as indicated in red. (B, C) Dual-luciferase reporter assays in THP-1 (B) and HL-60 (C) cells. miR-2110 mimics significantly reduced the activity of the wild-type ELK1 3′UTR reporter. Mutation of the predicted binding sites attenuated miR-2110-mediated repression to varying degrees, with MUT#1 showing the strongest loss of responsiveness, indicating that site 1 represents the dominant functional binding site within the ELK1 3′UTR. Firefly luciferase activity was normalized to Renilla luciferase activity and expressed relative to the corresponding mimic NC group. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined using two-tailed student’s t-test for comparisons between mimic NC and miR-2110 mimic groups within each reporter construct. ns, not significant; **P < 0.01; ***P < 0.001.

Article Snippet: miR-2110 mimics, negative control (NC) oligonucleotides, and ELK1 overexpression plasmids, along with their corresponding control vectors, were purchased from Sangon Biotech (Shanghai, China).

Techniques: Functional Assay, Binding Assay, Mutagenesis, Construct, Generated, Luciferase, Activity Assay, Two Tailed Test

Journal: Frontiers in Oncology

Article Title: miR-2110 orchestrates ERK–ELK1 transcriptional repression to induce cell-cycle arrest and enhance cytarabine sensitivity in acute myeloid leukemia

doi: 10.3389/fonc.2026.1833649

Figure Lengend Snippet: miR-2110 downregulates ELK1 and places ELK1 within a MAPK-associated interaction context. (A, B) qRT-PCR analysis showing reduced ELK1 mRNA levels in THP-1 and HL-60 cells after transfection with miR-2110 mimics. The NC group was used as the reference and set to 1.0. (C) Expression profile of ELK1 across multiple tumor and normal cohorts, showing relatively higher ELK1 expression in malignant tissues. (D) Protein–protein interaction network generated by querying the STRING database using ELK1 as the seed protein to identify known and predicted ELK1-associated interacting proteins in humans. (E) KEGG pathway enrichment analysis of ELK1-associated interacting proteins. (F) Gene Ontology molecular function enrichment analysis of ELK1-associated interacting proteins. Data in (A) and (B) are presented as mean ± SD from three independent experiments. Statistical significance was determined using two-tailed student’s t-test. ns, not significant; ***P < 0.001.

Article Snippet: miR-2110 mimics, negative control (NC) oligonucleotides, and ELK1 overexpression plasmids, along with their corresponding control vectors, were purchased from Sangon Biotech (Shanghai, China).

Techniques: Quantitative RT-PCR, Transfection, Expressing, Generated, Two Tailed Test

Journal: Frontiers in Oncology

Article Title: miR-2110 orchestrates ERK–ELK1 transcriptional repression to induce cell-cycle arrest and enhance cytarabine sensitivity in acute myeloid leukemia

doi: 10.3389/fonc.2026.1833649

Figure Lengend Snippet: ELK1 contributes to miR-2110–associated changes in cell-cycle-, apoptosis-, and MAPK-related protein expression in AML cells. (A) Western blot analysis and densitometric quantification of ELK1, cell-cycle–related proteins (Cyclin D1, CDK4, and p21), apoptosis-related proteins (Bcl-2, Bax, cleaved caspase-3, total caspase-3, cleaved PARP, and total PARP), and selected MAPK-associated markers (p-ERK1, total ERK1, c-FOS, and EGR1) in THP-1 cells transfected with negative control, miR-2110 mimics, ELK1 overexpression plasmid, or miR-2110 mimics combined with ELK1 overexpression. (B) Corresponding western blot analysis and quantification in HL-60 cells under the same experimental conditions. β-actin was used as the loading control. For cleaved proteins and phosphorylated proteins, band intensities were normalized to the corresponding total protein levels where applicable and then expressed as fold change relative to the NC group, which was set to 1.0. Other protein levels were normalized to β-actin and expressed relative to the NC group. (C, D) Immunofluorescence staining of ELK1 in THP-1 and HL-60 cells following the indicated treatments. Representative images and quantitative fluorescence intensity are shown. Scale bar = 100 μm. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined using one-way ANOVA followed by Tukey’s post hoc test. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: miR-2110 mimics, negative control (NC) oligonucleotides, and ELK1 overexpression plasmids, along with their corresponding control vectors, were purchased from Sangon Biotech (Shanghai, China).

Techniques: Expressing, Western Blot, Transfection, Negative Control, Over Expression, Plasmid Preparation, Control, Immunofluorescence, Staining, Fluorescence

Journal: Frontiers in Oncology

Article Title: miR-2110 orchestrates ERK–ELK1 transcriptional repression to induce cell-cycle arrest and enhance cytarabine sensitivity in acute myeloid leukemia

doi: 10.3389/fonc.2026.1833649

Figure Lengend Snippet: miR-2110 enhances Ara-C responsiveness and partially counteracts Ara-C resistance in AML cells. (A, B) Ara-C dose–response curves showing survival rates of parental THP-1 and HL-60 cells transfected with negative control, miR-2110 mimics, ELK1 overexpression plasmid, or miR-2110 mimics combined with ELK1 overexpression. (C, D) Ara-C dose–response curves in Ara-C–resistant THP-1 and HL-60 cells under the indicated transfection conditions. Cell survival was measured using CCK-8 assays and expressed relative to untreated cells in each corresponding group. (E, F) qRT-PCR analysis of endogenous ELK1 mRNA and miR-2110 expression in parental and Ara-C-resistant THP-1 cells (E) and HL-60 cells (F) . THP-1/Ara-C-R and HL-60/Ara-C-R indicate Ara-C-resistant THP-1 and HL-60 cells, respectively. Parental cells were used as the reference and set to 1.0. GAPDH was used as the internal control for ELK1 mRNA quantification, and U6 was used as the internal control for miR-2110 quantification. (G, H) Western blot analysis and densitometric quantification of ELK1, γH2AX, total H2AX, phosphorylated p53, total p53, cleaved caspase-3, total caspase-3, Bcl-2, and Bax in THP-1 and HL-60 cells treated with Ara-C. β-actin was used as the loading control. γH2AX and phosphorylated p53 were normalized to total H2AX and total p53, respectively; cleaved caspase-3 was normalized to total caspase-3; other proteins were normalized to β-actin. Values were expressed as fold change relative to the corresponding NC group, which was set to 1.0. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined using one-way ANOVA followed by Tukey’s post hoc test for multi-group comparisons. For dose–response curves, statistical comparisons were performed at the indicated concentrations. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: miR-2110 mimics, negative control (NC) oligonucleotides, and ELK1 overexpression plasmids, along with their corresponding control vectors, were purchased from Sangon Biotech (Shanghai, China).

Techniques: Transfection, Negative Control, Over Expression, Plasmid Preparation, CCK-8 Assay, Quantitative RT-PCR, Expressing, Control, Western Blot

Journal: Frontiers in Oncology

Article Title: miR-2110 orchestrates ERK–ELK1 transcriptional repression to induce cell-cycle arrest and enhance cytarabine sensitivity in acute myeloid leukemia

doi: 10.3389/fonc.2026.1833649

Figure Lengend Snippet: Graphical abstract illustrating the miR-2110–ELK1 regulatory axis in acute myeloid leukemia. miR-2110 is upregulated in pediatric AML and functions as a tumor suppressor by directly targeting the 3′UTR of ELK1, leading to reduced ELK1 expression and suppression of MAPK/ERK-associated transcriptional activity. This regulation results in inhibition of cell proliferation, induction of cell-cycle arrest, promotion of apoptosis, and increased sensitivity to Ara-C in AML cells. In vivo , miR-2110 overexpression suppresses tumor growth in a xenograft model. Overall, miR-2110–ELK1 signaling represents a potential therapeutic target in AML.

Article Snippet: miR-2110 mimics, negative control (NC) oligonucleotides, and ELK1 overexpression plasmids, along with their corresponding control vectors, were purchased from Sangon Biotech (Shanghai, China).

Techniques: Expressing, Activity Assay, Inhibition, In Vivo, Over Expression