Journal: Frontiers in Immunology

Article Title: Unveiling tumor senescence-driven prognostic heterogeneity via MALISS in stage II/III colorectal cancer

doi: 10.3389/fimmu.2025.1744719

Figure Lengend Snippet: NR1D2 promotes the CRC migration. (A) The importance of 30 immunosenescence-related genes selected by Coxboost algorithm. (B) Kaplan-Meier survival curve of PFS between patients with high NR1D2 and low NR1D2 mRNA level. (C) The NR1D2 expression of mRNA and protein levels within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3. Relative band intensity (normalized) is shown below the WB image. (D) SA-β-gal staining to analyze the senescence status in the HCT15 cells within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3. (E) P16 and P21 protein levels assessed by Western blot after NR1D2 knocking down. (F) Changes in mRNA expression of SASP-associated secretory factors (TGF-β, GDF15) as determined by qRT-PCR. (G) Wound healing assays was performed to detect the migration within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3 in HCT15. (H) Transwell assays was performed to detect the migration within native control, siNR1D2#1, siNR1D2#2 and siNR1D2#3 in HCT15. * means P < 0.05, ** means P < 0.01, *** means P < 0.001, **** means P < 0.0001.

Article Snippet: The HCT15 cell line (RRID: CVCL_0292), which was cultured in RPMI-1640 (Gibco) medium with 10% fetal bovine serum (Vazyme), penicillin-streptomycin (100 U/mL, NCM), was procured from ATCC.

Techniques: Migration, Expressing, Control, Staining, Western Blot, Quantitative RT-PCR

Journal: Cancer & Metabolism

Article Title: NSUN2 promotes colorectal cancer progression by stabilizing PHGDH mRNA to promote serine metabolism reprogramming

doi: 10.1186/s40170-025-00406-1

Figure Lengend Snippet: NSUN2 enhances the tumorigenesis and progression of CRC. A , B) After infection of SW620 (A) and HCT116 (B) cells with shRNAs, the expression of NSUN2 was analsyzed by Western blot (left) and qRT-PCR (right) assays. C) After infection of HCT15 cells with NSUN2-WT plasmids, the expression of NSUN2 was analyzed by Western blot (left) and qRT-PCR (right) assays. D-E) After knocking down NSUN2 in SW620 (D) and HCT116 (E) cells, cell proliferation ability was measured by the CCK8 assay. F) After knocking down NSUN2 in SW620 and HCT116 cells, cell colony formation ability was measured by the colony formation experiments. G) Cell proliferation ability was assessed using the CCK8 assay following NSUN2 overexpression in HCT15 cells. H) After overexpressing NSUN2 in HCT15 cells, cell proliferation ability was assessed by colony formation assay. I-J) After knocking down NSUN2 in SW620 (I) and HCT116 (J) cells, cell migration and invasion were determined by transwell assays. scale bars = 100 μm. K) After overexpressing NSUN2 in HCT115 cells, cell migration and invasion were determined by transwell assays. scale bars = 100 μm

Article Snippet: The human CRC cell lines HCT15, HCT116, SW620 were purchased from the American Type Culture Collection (ATCC).

Techniques: Infection, Expressing, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Over Expression, Colony Assay, Migration

Journal: Cancer & Metabolism

Article Title: NSUN2 promotes colorectal cancer progression by stabilizing PHGDH mRNA to promote serine metabolism reprogramming

doi: 10.1186/s40170-025-00406-1

Figure Lengend Snippet: NSUN2 promotes PHGDH expression in CRC. (A) Venn diagram of RNA-seq in NSUN2-stably knockdown HCT15 cells and combined it with MeRIP-seq data from the GEO public database ( GSE226129 ) to evaluate potential targets. Differential gene analysis between samples was carried out and screened by fold-change and P value. (B) PHGDH, PLEKHG2, VGF, LAMA5, KCTD15 and C6orf141 mRNA expression in SW620 cells and HCT116 cells with NSUN2 knockdown were detected by qRT-PCR. (C) Analyzing protein expression levels of PHGDH in SW620 and HCT116 cells after infection with shNSUN2-1 and shNSUN2-2. (D) Analyzing mRNA expression levels of PHGDH in SW620 and HCT116 cells after infection with shNSUN2-1 and shNSUN2-2. E , F) PHGDH was highly expressed in tumor tissues compared with adjacent normal tissues from GSE21510 (E) and TCGA (F) databases. G) Representative IHC staining images for PHGDH protein in human CRC specimens (scale bars = 250 μm and 50 μm), demonstrating its differential expression between tumor ( n = 56) and adjacent normal ( n = 56) tissue. H) Kaplan-Meier analysis of OS in the Renji Hospital cohort ( n = 180), comparing CRC patients with high ( n = 96) versus low ( n = 84) tumor PHGDH expression. I) NSUN2 were positively correlated with the expression of PHGDH at mRNA levels in TCGA databases. J) Tumor tissues and normal tissues from CRC patients were collected, and Western blot was used to analyze the levels of the NSUN2 and PHGDH proteins. K) Representative IHC staining for NSUN2 and PHGDH from CRC tissue (scale bars = 250 μm and 50 μm). Tumor 1# is representative of a patient with NSUN2-low CRC. Tumor 2# is representative of a patient with NSUN2-high CRC. L) Correlation of NSUN2 and PHGDH staining in human CRC tissue samples ( n = 180). NSUN2 and PHGDH show a positive correlation. ns, non-significant

Article Snippet: The human CRC cell lines HCT15, HCT116, SW620 were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, RNA Sequencing, Stable Transfection, Knockdown, Quantitative RT-PCR, Infection, Immunohistochemistry, Quantitative Proteomics, Western Blot, Staining

Journal: The Turkish Journal of Gastroenterology

Article Title: m6A Methylation Regulator RBM15-Mediated Upregulation of ITGBL1 mRNA Stability Aggravates Colon Adenocarcinoma Progression by Remodeling the Tumor Microenvironment

doi: 10.5152/tjg.2025.24068

Figure Lengend Snippet: Expression pattern of ITGBL1 in COAD. (A) The TIMER database was used to display the differential expression of ITGBL1 in the Cancer Genome Atlas (TCGA) pan-cancer dataset. (B) UALCAN database showed ITGBL1 expression in COAD based on sample types TCGA samples. (C and E) The UALCAN database shows the association between ITGBL1 expression and COAD individual cancer stage, nodal metastatic status, and histologic subtype. (F and G) KM plotter database was used to analyze the relationship between ITGBL1 expression and RFS and OS of COAD. (H) Immunohistochemical observation of ITGBL1 expression in normal and COAD tissues. (I) RT-qPCR assay was used to detect the expression level of ITGBL1 in 45 normal tissues and 46 COAD tumor tissues. (J and K) Western blot analysis of ITGBL1 protein level in normal tissues, COAD tumor tissues, NCM460 cell line, and COAD cell lines (DLD1, Lovo, HCT15, and SW620). ** P < .01, *** P < .001.

Article Snippet: Colon adenocarcinoma cell lines (DLD1, CL-0074; HCT15, CL-0097; Procell, Wuhan, China) were cultivated in corresponding media (CM-0074, CM-0097, Procell) at 37°C with 5% CO 2 .

Techniques: Expressing, Quantitative Proteomics, Immunohistochemical staining, Quantitative RT-PCR, Western Blot

Journal: The Turkish Journal of Gastroenterology

Article Title: m6A Methylation Regulator RBM15-Mediated Upregulation of ITGBL1 mRNA Stability Aggravates Colon Adenocarcinoma Progression by Remodeling the Tumor Microenvironment

doi: 10.5152/tjg.2025.24068

Figure Lengend Snippet: Effects of ITGBL1 downregulation in COAD cell proliferation, migration, invasion, and EMT. Lovo and SW620 cells were transfected with sh-NC, sh-ITGBL1#1, or sh-ITGBL1#2. (A) ITGBL1 protein level was determined using western blot in transfected Lovo and SW620 cells. (B and C) Cell viability and proliferation were measured in transfected Lovo and SW620 cells using CCK-8 and colony formation assays. (D and E) Cell migration and invasion were assessed using Transwell assays in transfected Lovo and SW620 cells. (F) Western blot analysis of E-cadherin, Vimentin, and N-cadherin protein levels in transfected Lovo and SW620 cells. *** P < .01.

Article Snippet: Colon adenocarcinoma cell lines (DLD1, CL-0074; HCT15, CL-0097; Procell, Wuhan, China) were cultivated in corresponding media (CM-0074, CM-0097, Procell) at 37°C with 5% CO 2 .

Techniques: Migration, Transfection, Western Blot, CCK-8 Assay

Journal: The Turkish Journal of Gastroenterology

Article Title: m6A Methylation Regulator RBM15-Mediated Upregulation of ITGBL1 mRNA Stability Aggravates Colon Adenocarcinoma Progression by Remodeling the Tumor Microenvironment

doi: 10.5152/tjg.2025.24068

Figure Lengend Snippet: Inhibiting ITGBL1 impaired COAD cell growthin vivo. SW620 cells stably infected with sh-NC or sh-ITGBL1 were respectively subcutaneously inoculated into mice. (A and B) Tumor growth curve of xenografts and representative images of the tumors at the end of the experiment were presented. (C) Tumor weight was measured. (D) IHC staining was performed to measure the positive expression of ITGBL1, Ki-67, N-cadherin, E-cadherin, and Vimentin in xenografts. ** P < .01, *** P < .001.

Article Snippet: Colon adenocarcinoma cell lines (DLD1, CL-0074; HCT15, CL-0097; Procell, Wuhan, China) were cultivated in corresponding media (CM-0074, CM-0097, Procell) at 37°C with 5% CO 2 .

Techniques: Stable Transfection, Infection, Immunohistochemistry, Expressing

Journal: The Turkish Journal of Gastroenterology

Article Title: m6A Methylation Regulator RBM15-Mediated Upregulation of ITGBL1 mRNA Stability Aggravates Colon Adenocarcinoma Progression by Remodeling the Tumor Microenvironment

doi: 10.5152/tjg.2025.24068

Figure Lengend Snippet: ITGBL1 regulates the immune response to cancer. (A) TIMER database displayed the correlation of ITGBL1 with tumor-infiltrating immune cells (CD8 + T cells, macrophages, CD4 + T cells, B cells, neutrophils, and myeloid dendritic cells) in COAD. (B) GEPIA database exhibited correlation of ITGBL1 with tumor-associated M2-type macrophage markers (MRC1, CD163, IL-10, and TGFB1) and PD-L1 (CD274) expression in COAD. (C) Expression association between ITGBL1 and PD-L1 in COAD tissues was evaluated using Pearson correlation analysis. (D-G) Lovo and SW620 cells were transfected with sh-NC or sh-ITGBL1. Then, CM of transfected Lovo and SW620 cells were co-culture with isolated macrophages (THP1-M0). (D) The proportion of CD206 + positive cells was measured using flow cytometry. (E and F) CD163, CD206, and IL-10 mRNA levels were detected using RT-qPCR. (G) CD163, CD206, and IL-10 protein levels were assessed using a western blot assay. (H and I) Effector CD8 + T cells were co-cultured with indicated target Lovo and SW620 cells. (H) CD8 + T cell proliferation was examined using CFSE staining. (I) CD8 + T cell apoptosis was monitored by flow cytometry assay. ** P < .01, *** P < .001.

Article Snippet: Colon adenocarcinoma cell lines (DLD1, CL-0074; HCT15, CL-0097; Procell, Wuhan, China) were cultivated in corresponding media (CM-0074, CM-0097, Procell) at 37°C with 5% CO 2 .

Techniques: Expressing, Transfection, Co-Culture Assay, Isolation, Flow Cytometry, Quantitative RT-PCR, Western Blot, Cell Culture, Staining

Journal: The Turkish Journal of Gastroenterology

Article Title: m6A Methylation Regulator RBM15-Mediated Upregulation of ITGBL1 mRNA Stability Aggravates Colon Adenocarcinoma Progression by Remodeling the Tumor Microenvironment

doi: 10.5152/tjg.2025.24068

Figure Lengend Snippet: RBM15 stabilizes ITGBL1 expression through methylation modification. (A) Changes in the m6A methylation level of ITGBL1 after inhibition of RBM15 were analyzed by MeRIP-qPCR assay. (B) Their interaction was verified using a dual-luciferase reporter assay in Lovo and SW620 cells. (C) ITGBL1 mRNA level was measured in Lovo and SW620 cells transfected with sh-NC, sh-RBM15, vector, or RBM15 using RT-qPCR. (D) RBM15 and ITGBL1 protein levels were assessed in Lovo and SW620 cells transfected with sh-NC, sh-RBM15, vector, or RBM15 using western blot. (E-H) The UALCAN database exhibited the association between RBM15 expression and COAD sample types, individual cancer stages, nodal metastatic status, and histologic subtypes. (I) The KM plotter database was applied to analyze the relationship between RBM15 expression and the OS of COAD. (J) IHC staining was used to detect the positive expression of RBM15 in normal and COAD tissues. (K) RBM15 mRNA level was determined in 45 normal tissues and 46 COAD tumor tissues using RT-qPCR. (L and M) Western blot analysis of RBM15 protein levels in normal tissues, COAD tumor tissues, NCM460 cell line, and COAD cell lines (Lovo and SW620). ** P < .01, *** P < .001.

Article Snippet: Colon adenocarcinoma cell lines (DLD1, CL-0074; HCT15, CL-0097; Procell, Wuhan, China) were cultivated in corresponding media (CM-0074, CM-0097, Procell) at 37°C with 5% CO 2 .

Techniques: Expressing, Methylation, Modification, Inhibition, Luciferase, Reporter Assay, Transfection, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: The Turkish Journal of Gastroenterology

Article Title: m6A Methylation Regulator RBM15-Mediated Upregulation of ITGBL1 mRNA Stability Aggravates Colon Adenocarcinoma Progression by Remodeling the Tumor Microenvironment

doi: 10.5152/tjg.2025.24068

Figure Lengend Snippet: RBM15/ITGBL1 regulated COAD cell proliferation, migration, invasion, and EMT. (A) The transfection efficiency of sh-RBM15#1 or sh-RBM15#2 in Lovo and SW620 cells was measured using western blot. (B-G) Lovo and SW620 cells were transfected with sh-NC, sh-RBM15#1, sh-RBM15#1+ vector, or sh-RBM15#1+ITGBL1. (B) Western blot analysis of RBM15 protein levels in transfected Lovo and SW620 cells. (C and D) CCK-8 and colony formation assays were performed to assess cell viability and proliferation. (E and F) Transwell assays were conducted to measure cell migration and invasion. (G) E-cadherin, Vimentin, and N-cadherin protein levels were determined using western blot. ** P < .01, *** P < .001.

Article Snippet: Colon adenocarcinoma cell lines (DLD1, CL-0074; HCT15, CL-0097; Procell, Wuhan, China) were cultivated in corresponding media (CM-0074, CM-0097, Procell) at 37°C with 5% CO 2 .

Techniques: Migration, Transfection, Western Blot, Plasmid Preparation, CCK-8 Assay

Journal: The Turkish Journal of Gastroenterology

Article Title: m6A Methylation Regulator RBM15-Mediated Upregulation of ITGBL1 mRNA Stability Aggravates Colon Adenocarcinoma Progression by Remodeling the Tumor Microenvironment

doi: 10.5152/tjg.2025.24068

Figure Lengend Snippet: m6A methylase RBM15-mediated upregulation of ITGBL1 mRNA stability could boost COAD cell proliferation, migration, invasion, EMT, and immune escape.

Article Snippet: Colon adenocarcinoma cell lines (DLD1, CL-0074; HCT15, CL-0097; Procell, Wuhan, China) were cultivated in corresponding media (CM-0074, CM-0097, Procell) at 37°C with 5% CO 2 .

Techniques: Migration