colorectal adenocarcinoma cell line hct15  (ATCC)

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