Fusobacterium nucleatum colonization is associated with decreased survival of helicobacter pylori-positive gastric cancer patients

Figure 1 Changes in the gene expression profile of 008L-C2 cancer cells cocultured with Fusobacterium nucleatum. Fusobacterium nucleatum was added to the cell culture at multiplicity of infection (MOI) 10 and 100. After 24 h and 72 h of incubation, the cells were rinsed extensively to remove unattached bacteria. RNA was collected and analyzed using RNA sequencing. The cells collected at time 0 were used as control. Sequencing reads were trimmed and mapped to hg19 using CLC Genomic Workbench v.12.0.3. The colored dots (green for MOI = 10 and red for MOI = 100) are genes showing more than four-fold change at the respective incubation time. Genes with transcripts per million < 1 in all datasets and less than a four-fold change are not shown.

Figure 2 Identification and ontological analysis of the RNA sequencing dataset. A: The expression dynamics of the interferon response, cytokines, and chemokines genes are shown; B: The expression dynamic of the antiviral genes is shown; C: The expression dynamics of actin, its regulators, and genes involved in cell adhesion are shown. MOI: Multiplicity of infection; TPM: Transcripts per million.

Figure 3 Identification of Fusobacterium nucleatum in the resected gastric cancer tissues. A: Fusobacterium nucleatum (F. nucleatum) was detected by amplification of the conserved NusG gene sequence using nested polymerase chain reaction method. The end point product was analyzed using 6% nondenaturing polyacrylamide gel electrophoresis; B: The relative risk of F. nucleatum colonization was analyzed against the sex, age, status of Helicobacter pylori colonization, and cancer stage. Data analysis is shown as Forest plot.

Figure 4 Analysis of Fusobacterium nucleatum colonization in gastric cancer patients. A: The study cohort was divided into Helicobacter pylori (H. pylori)-positive and negative groups according to the result of nested-polymerase chain reaction analysis. Survival probability was calculated using Kaplan-Meier analysis in all or stage II, III, and IV cancer patients with at least 1-year follow-up; B: Survival probability was calculated in stages II, III, and IV cancer patients.