Journal: Scientific Reports

Article Title: Single-cell and bulk transcriptomics reveal Fusobacterium nucleatum promotes colorectal cancer metastasis by upregulating LAMC2 in in vitro and in vivo models

doi: 10.1038/s41598-025-26249-w

Figure Lengend Snippet: F. nucleatum promotes the migration of CRC cells in vitro. ( A ) Scanning electron microscopy and ( B ) transmission electron microscopy images show rod-shaped bacteria (arrows) penetrating and invading HCT116 cells. ( C - J ) LOVO and HCT116 cells were incubation with PBS control, E.coli , or F. nucleatum . Cell migration ability was evaluated through wound healing assays and Transwell assays ( n = 4 per group, One-way ANOVA test). In the Transwell assays, migrated cells were stained with crystal violet, and images were randomly captured under a microscope, with 5 random fields per sample counted. Statistical analysis was performed using one-way ANOVA. Data are shown as mean ± SD.* represents p < 0 0.05, ** represents p < 0 0.01, *** represents p < 0.001, and **** represents p < 0.0001, ns represents no statistical significant.

Article Snippet: Human CRC cell lines (HCT116, LOVO, HCT8, SW480 and SW620) were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Migration, In Vitro, Electron Microscopy, Transmission Assay, Bacteria, Incubation, Control, Staining, Microscopy

Journal: Scientific Reports

Article Title: Single-cell and bulk transcriptomics reveal Fusobacterium nucleatum promotes colorectal cancer metastasis by upregulating LAMC2 in in vitro and in vivo models

doi: 10.1038/s41598-025-26249-w

Figure Lengend Snippet: F. nucleatum upregulates LAMC2 in CRC cells. ( A ) Volcano plot showing the expression profile of differentially expressed genes (DEGs) in LOVO cells between F. nucleatum infection and PBS control treatment ( n = 3 per group, p < 0.05 and |log2 (fold change)| ≥ 1). ( B ) Intersection genes between the up DEGs from the GSE173549 dataset and the metastasis-related genes from the CancerSEA database. ( C and D ) The expression of LAMC2 was analyzed by single-cell RNA sequencing data GSE132465 . The cells were classified into seven distinct clusters, including T, B, myeloid, mast, epithelial, endothelial, and mesenchymal cells. The statistical significance between two groups was evaluated by wilcox.test. Normal: adjacent normal tissues of CRC, Tumor: tumor tissues of CRC. ( E and F )The mRNA and protein expression of LAMC2 after F. nucleatum or PBS treatment in CRC cells ( n = 4 per group, One-way ANOVA test). ( G and H ) The mRNA expression of LAMC2 after F. nucleatum , E. coli or PBS treatment in CRC cells ( n = 4 per group, One-way ANOVA test). ( I and K ) The protein expression of LAMC2 after PBS treatment, F. nucleatum infection of MOI = 100 or 500, or E. coli infection of MOI = 100 in CRC cells (repeated three times). ( J and L ) The protein expression of LAMC2 at 0, 4, 12, 24 and 48 h after F. nucleatum infection in CRC cells ( n = 4 per group, One-way ANOVA test). Data are shown as mean ± SD.* represents p < 0 0.05, ** represents p < 0 0.01, *** represents p < 0.001, and **** represents p < 0.0001, ns represents no statistical significant.

Article Snippet: Human CRC cell lines (HCT116, LOVO, HCT8, SW480 and SW620) were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Infection, Control, RNA Sequencing

Journal: Scientific Reports

Article Title: Single-cell and bulk transcriptomics reveal Fusobacterium nucleatum promotes colorectal cancer metastasis by upregulating LAMC2 in in vitro and in vivo models

doi: 10.1038/s41598-025-26249-w

Figure Lengend Snippet: F. nucleatum promotes metastasis of CRC cells in a LAMC2 dependent manner. ( A and B ) Detection of the knockdown efficiency of LAMC2 by different sequences of lentiviral vectors at the mRNA ( n = 4 per group, One-way ANOVA test) and protein (repeated three times) levels. LOVO cell were transfected with LAMC2 shRNA lentivirus or control lentivirus, and then incubated with F. nucleatum or PBS. ( C and D ) The mRNA expression and protein expression of LAMC2 were measured by qPCR ( n = 4, One-way ANOVA test) and western blot (repeated three times). ( E and G ) The motility was detected by wound-healing assay ( n = 6, One-way ANOVA test) or ( F and H ) transwell assay( n = 5, One-way ANOVA test). ( I ) LOVO cells constructed with lentivirus were incubated with F. nucleatum or PBS and then injected into the tail vein of nude mice (5 mice per group). ( J and K ) HE staining represents the images and the number of metastatic lesions (Student’s t-test) in each group. Data are shown as mean ± SD. * represents p < 0 0.05, ** represents p < 0 0.01, *** represents p < 0.001, and **** represents p < 0.0001, ns represents no statistical significant.

Article Snippet: Human CRC cell lines (HCT116, LOVO, HCT8, SW480 and SW620) were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Knockdown, Transfection, shRNA, Control, Incubation, Expressing, Western Blot, Wound Healing Assay, Transwell Assay, Construct, Injection, Staining

Journal: Scientific Reports

Article Title: Single-cell and bulk transcriptomics reveal Fusobacterium nucleatum promotes colorectal cancer metastasis by upregulating LAMC2 in in vitro and in vivo models

doi: 10.1038/s41598-025-26249-w

Figure Lengend Snippet: F.nucleatum upregulates the expression of LAMC2 through hsa-miRNA-7977. ( A ) western blot (repeated three times) was used to detect the overexpression efficiency of LAMC2 . HCT116 cell were transfected with LAMC2 oe-RNA lentivirus or control lentivirus, and then incubated with F. nucleatum or PBS. ( B and C ) The protein expression and mRNA expression of LAMC2 were measured by western blot (repeated three times) and qPCR ( n = 4, One-way ANOVA test). ( D and F ) The motility was detected by wound-healing assay ( n = 6, One-way ANOVA test) or ( E and G ) transwell assay ( n = 5, One-way ANOVA test). ( H ) Prediction of miRNA binding to LAMC2 using public databases, with a Venn diagram showing overlapping genes from the MiRDB, MirDIP, and Targetscan databases. ( I ) A Venn diagram showing candidate miRNAs binding to LAMC2 predicted by three public databases (miRNA- LAMC2 ) and downregulated miRNAs after F. nucleatum infection in CRC cells (Fn-miRNA). ( J ) Detection of miR-7977 expression using qPCR in CRC cells after F. nucleatum infection ( n = 4, One-way ANOVA test). ( K )Prediction of the binding site of has-miR-7977 to the 3’-UTR of LAMC2 using the Targetscan database.

Article Snippet: Human CRC cell lines (HCT116, LOVO, HCT8, SW480 and SW620) were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Western Blot, Over Expression, Transfection, Control, Incubation, Wound Healing Assay, Transwell Assay, Binding Assay, Infection