IADR Abstract Archives
Metabolic Gene Expression Profiles (Transcriptomics) by Oral Carcinoma Cells Upon Stimulation With Porphyromonas Gingivalis
Objectives:
Periodontitis, a chronic inflammatory disease, is caused by an infectious biofilm, affecting all periodontal components. It has been implicated as a cause of cancerous lesions, as chronic inflammation intensifies the risk for carcinomas. Interactions between periodontal pathogens and the host immune response initiate periodontitis and are responsible for its subsequent progression. Porphyromonas gingivalis (P. gingivalis), a Gram-negative anaerobic rod that expresses various virulence factors, is considered as cornerstone pathogen in periodontal biofilms. The aim of this study was to investigate the impact of P. gingivalis W83 membranes on RNA expression of oral squamous carcinoma cells using transcriptomic analysis.
Methods:
Human squamous cell carcinoma cells (SCC-25) were incubated for 4h and 24h respectively with the membrane fraction from P. gingivalis., following which cells were harvested and RNA extracted. RNA sequencing was performed and differential gene expression and enrichment was analyzed using the GO, KEGG and REACTOME software suites. Results of the transcriptome analysis was validated using quantitative real time PCR (RTPCR) of selected genes.
Results:
Differential gene expression analysis yielded 15 genes, which were up-regulated and 1 down-regulated after 4h and 61 up-regulated and 278 down-regulated after 24h. GO, KEGG and REACTOME enrichment analysis revealed a strong metabolic transcriptomic response signature. At 4h/24h post treatment, there were significant alteration in genes involved in metabolic pathways and replication. RT-PCR of selected genes belonging to immune response signaling and drug metabolism confirmed up-regulation of CCL20, CXCL8, NFkBIA, TNFAIP3, TRAF5, CYP1A1 and NOD2.
Conclusions:
This work sheds light on the RNA transcriptome of human oral squamous carcinoma cells following stimulation with P. gingivalis membranes and identifies a strong immune and metabolic gene expression response to components in the envelope of this periodontal pathogen. The data provide a basis for further integrated studies of molecular and cellular interactions between P. gingivalis and oral epithelium to elucidate basic mechanisms of periodontitis and the development of cancer.
This study was supported by a grant of the German-Research-Foundation
(DFG, GR-3365/4-1).
Periodontitis, a chronic inflammatory disease, is caused by an infectious biofilm, affecting all periodontal components. It has been implicated as a cause of cancerous lesions, as chronic inflammation intensifies the risk for carcinomas. Interactions between periodontal pathogens and the host immune response initiate periodontitis and are responsible for its subsequent progression. Porphyromonas gingivalis (P. gingivalis), a Gram-negative anaerobic rod that expresses various virulence factors, is considered as cornerstone pathogen in periodontal biofilms. The aim of this study was to investigate the impact of P. gingivalis W83 membranes on RNA expression of oral squamous carcinoma cells using transcriptomic analysis.
Methods:
Human squamous cell carcinoma cells (SCC-25) were incubated for 4h and 24h respectively with the membrane fraction from P. gingivalis., following which cells were harvested and RNA extracted. RNA sequencing was performed and differential gene expression and enrichment was analyzed using the GO, KEGG and REACTOME software suites. Results of the transcriptome analysis was validated using quantitative real time PCR (RTPCR) of selected genes.
Results:
Differential gene expression analysis yielded 15 genes, which were up-regulated and 1 down-regulated after 4h and 61 up-regulated and 278 down-regulated after 24h. GO, KEGG and REACTOME enrichment analysis revealed a strong metabolic transcriptomic response signature. At 4h/24h post treatment, there were significant alteration in genes involved in metabolic pathways and replication. RT-PCR of selected genes belonging to immune response signaling and drug metabolism confirmed up-regulation of CCL20, CXCL8, NFkBIA, TNFAIP3, TRAF5, CYP1A1 and NOD2.
Conclusions:
This work sheds light on the RNA transcriptome of human oral squamous carcinoma cells following stimulation with P. gingivalis membranes and identifies a strong immune and metabolic gene expression response to components in the envelope of this periodontal pathogen. The data provide a basis for further integrated studies of molecular and cellular interactions between P. gingivalis and oral epithelium to elucidate basic mechanisms of periodontitis and the development of cancer.
This study was supported by a grant of the German-Research-Foundation
(DFG, GR-3365/4-1).