Journal: Frontiers in Cell and Developmental Biology

Article Title: ALYREF regulates glycolysis and promotes the progression of endometrial cancer through m 5 C modification of PKM2 mRNA

doi: 10.3389/fcell.2026.1831562

Figure Lengend Snippet: ALYREF promotes proliferation, migration, and invasion in endometrial cancer cells. (A) Western blot analysis of ALYREF protein expression in normal endometrial cell line (EEC) and EC cell lines (Ishikawa, HEC-1-A, AN3CA). (B) Validation of ALYREF knockdown and overexpression by Western blot and RT-qPCR. (C) CCK-8 assay showing cell viability after ALYREF modulation. (D) Colony formation assays evaluating the proliferative ability of Ishikawa and HEC-1-A cells with altered ALYREF expression. (E,F) Transwell migration and invasion assays in Ishikawa (E) and HEC-1-A (F) cells with ALYREF knockdown or overexpression; Scale bar: 50 μm. (G) Western blot and quantification of EMT-related markers (E-cadherin, N-cadherin, Vimentin, MMP2). sh-NC: scramble shRNA negative control for knockdown; Vector: empty vector control for overexpression. Data are presented as mean ± SD from three independent biological replicates; two-tailed unpaired t -test; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies included ALYREF (Proteintech, 16690-1-AP), PKM2 (Proteintech, 15822-1-AP), E-cadherin (Proteintech, 20874-1-AP), N-cadherin (Proteintech, 22018-1-AP), Vimentin (Proteintech, 10366-1-AP), MMP2 (Proteintech, 10373-2-AP), and β-actin (Proteintech, 66009-1-Ig).

Techniques: Migration, Western Blot, Analysis, Expressing, Biomarker Discovery, Knockdown, Over Expression, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, shRNA, Negative Control, Plasmid Preparation, Control, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: ALYREF regulates glycolysis and promotes the progression of endometrial cancer through m 5 C modification of PKM2 mRNA

doi: 10.3389/fcell.2026.1831562

Figure Lengend Snippet: ALYREF promotes tumor growth in vivo and distant metastasis in endometrial cancer. (A) Representative images of xenograft tumors derived from Ishikawa cells with or without ALYREF knockdown. (B) Tumor volume growth curves over time. (C) Tumor weight at endpoint. (D) Hematoxylin-eosin (HE) staining of xenograft tumors. (E) IHC staining and quantification of ALYREF and Ki-67 expression in tumor tissues. (F) Representative gross images of lung and liver tissues from tail-vein injected mice (sh-NC vs. sh-ALYREF) and quantification of metastatic nodules. (G) HE staining of lung and liver tissues to evaluate metastatic lesions. (H) IHC staining and scoring of E-cadherin and Vimentin in metastatic lesions of the lung and liver. For tumor formation (A–C) and metastasis (F) , five mice per group were used; HE and IHC analyses were performed on three mice per group. Tumor growth curves (B) were analyzed by two-way repeated-measures ANOVA with Bonferroni post hoc test; other comparisons were made by two-tailed unpaired t -test. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies included ALYREF (Proteintech, 16690-1-AP), PKM2 (Proteintech, 15822-1-AP), E-cadherin (Proteintech, 20874-1-AP), N-cadherin (Proteintech, 22018-1-AP), Vimentin (Proteintech, 10366-1-AP), MMP2 (Proteintech, 10373-2-AP), and β-actin (Proteintech, 66009-1-Ig).

Techniques: In Vivo, Xenograft Assay, Derivative Assay, Knockdown, H&E Stain, Immunohistochemistry, Expressing, Injection, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: ALYREF regulates glycolysis and promotes the progression of endometrial cancer through m 5 C modification of PKM2 mRNA

doi: 10.3389/fcell.2026.1831562

Figure Lengend Snippet: PKM2 contributes to the oncogenic effects of ALYREF in endometrial cancer cells (A) Validation of PKM2 knockdown efficiency at the protein and mRNA levels by Western blot and RT-qPCR, respectively. (B,C) Cell viability (B) and colony formation (C) assays demonstrating suppressed cell proliferation after PKM2 knockdown. (D,E) Transwell assays showing reduced cell migration (D) and invasion (E) abilities after PKM2 knockdown. (F–I) Functional rescue assays illustrating the effects of PKM2 overexpression on cell viability (F) , colony formation (G) , migration (H) , and invasion (I) in the indicated groups (Con, ov-PKM2, sh-ALYREF, and sh-ALYREF + ov-PKM2). (J) Western blot analysis and corresponding quantification of epithelial-mesenchymal transition (EMT) markers (E-cadherin, N-cadherin, Vimentin, and MMP2) across the indicated groups (Con, ov-PKM2, sh-ALYREF, and sh-ALYREF + ov-PKM2). β-actin was used as a loading control. All data are presented as mean ± SD from three independent biological replicates. (A–E) Two-tailed unpaired t -test; (F–J) One-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies included ALYREF (Proteintech, 16690-1-AP), PKM2 (Proteintech, 15822-1-AP), E-cadherin (Proteintech, 20874-1-AP), N-cadherin (Proteintech, 22018-1-AP), Vimentin (Proteintech, 10366-1-AP), MMP2 (Proteintech, 10373-2-AP), and β-actin (Proteintech, 66009-1-Ig).

Techniques: Biomarker Discovery, Knockdown, Western Blot, Quantitative RT-PCR, Colony Assay, Migration, Functional Assay, Over Expression, Analysis, Control, Two Tailed Test