Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: MMP-9 , a factor that promotes Vasculogenic mimicry, is highly expressed in CC and is associated with poor prognosis. (A) CC database of TCGA was used to analyze key factors associated with VM. (B) Association of Sox2 expression with overall survival in CC (log-rank test). (C) Association of MMP-9 expression with overall survival in CC (log-rank test). (D) Panoramic scans after immunohistochemical detection of MMP-9 and H&E staining in samples from cancerous and paracancerous tissues from subjects with CC. Scale bar, 50 µm. Original magnification, ×20. (E) Protein levels of MMP-9 in 20 paired samples, with the MMP-9 level in CC tissue expressed compared with that in the paired normal tissue. (F) Expression levels of MMP-9 mRNA in 44 paired CC and paracancerous tissues, with MMP-9 expression in CC tissue expressed compared with that in the paired normal tissue. (G) Comparison of the average expression levels of MMP-9 mRNA in CC tissues compared with paracancerous tissues. (H) HeLa and SiHa cells were incubated under hypoxia (0.1% O 2 ) and proteins collected at 24, 48 and 72 h for western blotting of ALDH1, EPHA2, MMP-9 and GAPDH. ImageJ was used to semi-quantify western blotting signals from HeLa (I) and SiHa (J) cells. GAPDH served as an internal reference. *P<0.05, **P<0.01 and ***P<0.001. MMP-9, matrix metalloproteinase 9; VM, vasculogenic mimicry; ALDH1, aldehyde dehydrogenase 1; EPHA2, ephrin type-A receptor 2; TCGA, The Cancer Genome Atlas; Sox2, SRY-box transcription factor 2; CC, cervical cancer; CESC, cervical squamous cell carcinoma.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Immunohistochemical staining, Staining, Comparison, Incubation, Western Blot

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 promotes Vasculogenic mimicry, invasion and migration of CC cells under hypoxic conditions. (A) Expression levels of NSUN2 in CC cells lines (HeLa, SiHa, CaSki, C33A and HT-3) and in a normal cervical cell line (HaCaT). (B) RT-qPCR was used to determine relative expression levels of NSUN2 mRNA in HeLa and SiHa cells after transfection of shRNAs and incubation under hypoxia for 24 h. (C) Relative expression levels of MMP-9 mRNA in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 24 h. (D) Dot blot assay analysis of m 5 C expression levels in HeLa and SiHa cells after 48 h of hypoxia culture following transfection with NSUN2 knockdown plasmids. (E) Semi-quantitative analysis of dot blot results in HeLa cells. (F) Semi-quantitative analysis of dot blot results in SiHa cells. (G) Western blotting was used to investigate NSUN2 and MMP-9 protein levels in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 48 h. ImageJ was used to semi-quantify western blotting bands for (H) NSUN2 and (I) MMP-9 protein levels. (J) 2D tube-forming assays of HeLa and SiHa cells transfected with an NSUN2 -interfering plasmid and incubated under hypoxia. Scale bar, 100 µm. Original magnification, ×4. (K) Assay displayed in panel (J) was quantified using Image Pro. (L) Invasion assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (M) Assay displayed in panel was quantified using Image Pro. (N) Migration assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (O) Assay displayed in panel (N) was quantified using Image Pro. *P<0.05, **P<0.01 and ***P<0.001. VM, Vasculogenic mimicry; CC, cervical cancer; IHC, immunohistochemistry; NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; RT-qPCR, reverse transcription-quantitative PCR; IHC, immunohistochemistry; CaSki, human cervical cancer cell line with intestinal metastasis; C33A, human cervical cancer cell line; HaCaT, human skin keratinocytes cell line; HeLa, human cervical cancer cell line; HT-3, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Migration, Expressing, Quantitative RT-PCR, Transfection, Incubation, Plasmid Preparation, Dot Blot, Knockdown, Quantitative Dot Blot, Western Blot, Invasion Assay, Control, Immunohistochemistry, Reverse Transcription, Real-time Polymerase Chain Reaction, Cloning, shRNA, Negative Control

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 increases the stability of MMP-9 mRNA. (A) A positive correlation was observed between NSUN2 and MMP-9 mRNA expression levels in 44 pairs of samples from subjects with CC. Enrichment of the m 5 C modification of MMP-9 mRNA in HeLa (B) and SiHa (C) Cells were measured with anti-m 5 C methylated-RNA IP assays. Interaction of NSUN2 with MMP-9 mRNA in (D) HeLa and (E) SiHa cells was measured with anti- NSUN2 RNA IP assays. Stability of MMP-9 mRNA after interference with and overexpression of NSUN2 was measured in (F) HeLa and (G) SiHa cells. (H) A model illustrating the proposed mechanism by which NSUN2-mediated stabilization of MMP-9 mRNA promotes Vasculogenic mimicry in CC. NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; HeLa, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; IP, immunoprecipitation; CC, cervical cancer; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Modification, Methylation, Over Expression, Plasmid Preparation, Cloning, shRNA, Immunoprecipitation, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Maresin 1 activates autophagy through SIRT1/PPAR-γ signaling to mitigate high glucose-induced pyroptosis in human retinal pigment epithelial cells

doi: 10.3892/etm.2026.13134

Figure Lengend Snippet: MaR1 ameliorates HG-induced pyroptosis in ARPE-19 cells. (A) Effect of MaR1 treatment at varying concentrations on the fluorescence intensity of GSDMD-N in HG-induced ARPE-19 cells. Scale bar, 50 µm. (B) Effect of MaR1 treatment at varying concentrations on the expression of pyroptosis-related proteins (caspase-1, GSDMD, GSDMD-N, cleaved caspase-1, NLRP3, ASC and IL-18) in HG-induced ARPE-19 cells. * P<0.05, ** P<0.01 and *** P<0.001. MaR1, maresin 1; HG, high glucose; NG, normal glucose; GSDMD, gasdermin D; GSDMD-N, GSDMD N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD.

Article Snippet: Subsequently, the cells were blocked with 1% BSA solution (cat. no. ST023; Beyotime Biotechnology) at room temperature for 1 h, followed by overnight incubation at 4 ̊C with diluted anti-GSDMD N-terminal (GSDMD-N) primary antibody (1:100; cat. no. ER1901-37; HUABIO).

Techniques: Fluorescence, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Maresin 1 activates autophagy through SIRT1/PPAR-γ signaling to mitigate high glucose-induced pyroptosis in human retinal pigment epithelial cells

doi: 10.3892/etm.2026.13134

Figure Lengend Snippet: MaR1 mediates autophagy through SIRT1/peroxisome proliferator-activated receptor-γ signaling to ameliorate HG-induced pyroptosis in ARPE-19 cells. (A) SIRT1 inhibitor EX527 and autophagy inhibitor 3-MA enhanced the fluorescence intensity of GSDMD-N in ARPE-19 cells treated with HG and MaR1. Scale bar, 50 µm. (B) EX527 and 3-MA upregulated the expression of GSDMD-N, cleaved caspase-1, NLRP3, ASC and IL-18 in ARPE-19 cells treated with HG and MaR1. * P<0.05, ** P<0.01 and *** P<0.001. MaR1, maresin 1; HG, high glucose; NG, normal glucose; SIRT1, sirtuin 1; GSDMD, gasdermin D; GSDMD-N, GSDMD N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD; 3-MA, 3-methyladenine.

Article Snippet: Subsequently, the cells were blocked with 1% BSA solution (cat. no. ST023; Beyotime Biotechnology) at room temperature for 1 h, followed by overnight incubation at 4 ̊C with diluted anti-GSDMD N-terminal (GSDMD-N) primary antibody (1:100; cat. no. ER1901-37; HUABIO).

Techniques: Fluorescence, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Maresin 1 activates autophagy through SIRT1/PPAR-γ signaling to mitigate high glucose-induced pyroptosis in human retinal pigment epithelial cells

doi: 10.3892/etm.2026.13134

Figure Lengend Snippet: Schematic illustration of the protective mechanism of MaR1 against HG-induced pyroptosis in ARPE-19 cells. High glucose stimulation triggers the overproduction of ROS, leading to the activation of the NLRP3 inflammasome complex (comprising NLRP3, ASC and Caspase-1). This activation results in the cleavage of GSDMD, which forms membrane pores (GSDMD-N) and promotes the release of pro-inflammatory cytokines, ultimately inducing pyroptosis. However, MaR1 treatment effectively upregulates SIRT1 expression, which in turn enhances PPAR-γ levels and restores autophagic flux. The restored autophagy facilitates the clearance of damaged mitochondria and ROS, thereby inhibiting the NLRP3/GSDMD signaling pathway and protecting ARPE-19 cells from HG-induced inflammatory damage. SIRT1, sirtuin 1; PPAR-γ, peroxisome proliferator-activated receptor-γ; ROS, reactive oxygen species; GSDMD-N, gasdermin D N-terminal; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD.

Article Snippet: Subsequently, the cells were blocked with 1% BSA solution (cat. no. ST023; Beyotime Biotechnology) at room temperature for 1 h, followed by overnight incubation at 4 ̊C with diluted anti-GSDMD N-terminal (GSDMD-N) primary antibody (1:100; cat. no. ER1901-37; HUABIO).

Techniques: Activation Assay, Membrane, Expressing

Journal: BioMed Research International

Article Title: The Effects of Neonatal Zingerone Administration and Adolescent Alcohol Exposure on Bone Health Markers and Morphometry in Male Sprague–Dawley Rats

doi: 10.1155/bmri/7060593

Figure Lengend Snippet: Effects of neonatally administered zingerone (40 mg/kg/day) and adolescent alcohol (1 g/kg) on plasma concentrations of (A) BALP, (B) OC, and (C) P1NP as bone health markers. Data were presented as mean ± standard deviation. BALP, OC, and P1NP levels were elevated in the alcohol‐treated groups with or without zingerone. Statistically significant if ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001 between treatment groups. Group 1 ( C ) received no treatment in the neonatal phase and during adolescence; Group 2 ( Z + W ) was treated with zingerone (40 mg/kg) in the neonatal phase and water during adolescence; Group 3 ( A + A ) treated with alcohol during both the neonatal and adolescent phase (1 g/kg ethanol); Group 4 ( Z + A ) treated with zingerone in the neonatal phase and alcohol in adolescence; Group 5 (ZA + A ) treated with a combination of zingerone–alcohol in the neonatal phase, followed by alcohol in adolescence. A —alcohol; C —control; W —water; Z —zingerone; ZA—zingerone‐alcohol; BALP—bone‐specific alkaline phosphatase, OC—osteocalcin, P1NP—procollagen Type 1 N‐terminal propeptide.

Article Snippet: The plasma activity of BALP (Catalog No. ER0761, FineTest, Wuhan, China), OC (Catalog No. ER1205, FineTest, Wuhan, China), and P1NP (Catalog No. E‐EL‐R1414, Elabscience, Houston, Texas, United States) were determined in rat plasma as described by the manufacturer′s instructions.

Techniques: Clinical Proteomics, Standard Deviation, Control