IADR Abstract Archives
Effects of Sphingosine-1-Phosphate Receptor 2 (S1PR2) on the Behaviour of Three Oral Squamous Cell Carcinoma (OSCC) Lines.
Objectives: This study aimed to determine the effects of S1PR2 on proliferation, migration and invasion in three OSCC lines.
Methods: H357, H400 and H413 cell lines were treated with 10µM JTE013 (S1PR2 antagonist) and CYM5478 (S1PR2 agonist). Relative expression levels of S1PR1-5 genes were determined using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Cell counts and the BrdU assay were used to determine cell growth and proliferation in response to treatments. Scratch-wound and transwell migration assays were used to examine the influence of the treatments on migration and invasion. Statistical analysis of the differences observed between experiments was undertaken using a one-way ANOVA.
Results: RT-PCR revealed that the three cell lines expressed all five S1PR subtypes. Inhibition of S1PR2 by JTE013 treatment caused a significant decrease in cell proliferation: the growth curves revealed an increase in the doubling time, whilst the BrdU assay also showed a decrease in the proportion of BrdU+ cells. JTE013 treatment also significantly reduced migration and decreased invasion (p<0.05) in all three cell lines. In contrast, stimulation of S1PR2 by CYM5478 did not significantly increase cell proliferation and resulted in varying effects on migration, depending on the cell line and experimental assays used. H400 cells showed an increased in migration (scratch-wound assay, p<0.01), whilst the transwell migration assay revealed increased migration in H357 and H413 cells (p<0.05). S1PR2 stimulation increased invasion (p<0.01) in all cell lines.
Conclusions: S1PR2 influences the proliferation, migration and invasion of the three OSCC cell lines, but its effects are cell-line specific. The results suggest S1PR2 as a potential target for OSCC treatment development and provide a further start point to unravel the pathways that dictate tumour proliferation, migration and invasion in OSCC. Migration variability after SP1R2 stimulation depending on assay types used suggests that tumour cell migration is multi-factorial process.
Methods: H357, H400 and H413 cell lines were treated with 10µM JTE013 (S1PR2 antagonist) and CYM5478 (S1PR2 agonist). Relative expression levels of S1PR1-5 genes were determined using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Cell counts and the BrdU assay were used to determine cell growth and proliferation in response to treatments. Scratch-wound and transwell migration assays were used to examine the influence of the treatments on migration and invasion. Statistical analysis of the differences observed between experiments was undertaken using a one-way ANOVA.
Results: RT-PCR revealed that the three cell lines expressed all five S1PR subtypes. Inhibition of S1PR2 by JTE013 treatment caused a significant decrease in cell proliferation: the growth curves revealed an increase in the doubling time, whilst the BrdU assay also showed a decrease in the proportion of BrdU+ cells. JTE013 treatment also significantly reduced migration and decreased invasion (p<0.05) in all three cell lines. In contrast, stimulation of S1PR2 by CYM5478 did not significantly increase cell proliferation and resulted in varying effects on migration, depending on the cell line and experimental assays used. H400 cells showed an increased in migration (scratch-wound assay, p<0.01), whilst the transwell migration assay revealed increased migration in H357 and H413 cells (p<0.05). S1PR2 stimulation increased invasion (p<0.01) in all cell lines.
Conclusions: S1PR2 influences the proliferation, migration and invasion of the three OSCC cell lines, but its effects are cell-line specific. The results suggest S1PR2 as a potential target for OSCC treatment development and provide a further start point to unravel the pathways that dictate tumour proliferation, migration and invasion in OSCC. Migration variability after SP1R2 stimulation depending on assay types used suggests that tumour cell migration is multi-factorial process.