Abstract: Objective To investigate the impact of Capnocytophaga leadbetteri (C. leadbetteri ), isolated from tissues of patients with tongue squamous cell carcinoma (TSCC), on the proliferation, invasion, and migration of TSCC Cal27 cell line. Methods The C. leadbetteri strain n95, previously isolated and cultured by our research group from the tumor tissue of TSCC patients, were identified through revival culture, colony morphology observation, Gram staining, 16S rRNA sequencing analysis, and phylogenetic tree construction. Cal27 cells were infected with C. leadbetteri n95 suspension (CAL27⁃C. L group), while uninfected Cal27 cells (CAL27 group) served as the control. Adhesion counts of cells and bacterial were recorded 2 h post⁃infection. The proliferative capacity of Cal27 cells at different multiplicity of infection (MOI=0, 10, 25) and time points (0, 24, 48, 72 h) was assessed using the CCK⁃8 assay. Transwell assays were employed to evaluate the migration and invasion capabilities of cells. Results Based on 16S rRNA sequences alignment, which exhibits a 99.63% sequence identity with the reference strain H6253, and Neighbour⁃Joining phylogenetic tree, strain n95 was identified as C. leadbetteri. Cell adhesion assays demonstrated that the adhesion index of C. leadbetteri n95 to Cal27 cells was 1.62±0.57. The CCK⁃8 assay results demonstrated that the proliferation activity of Cal27 cells was significantly enhanced  (P<0.05) after 24 h of infection with C. leadbetteri n95 at an MOI of 10∶1 . Transwell assays revealed that the invasion and migration capabilities of the CAL27⁃C. L group were significantly enhanced compared to the CAL27 group (P<0.01). Conclusions C. leadbetteri n95, isolated from TSCC, effectively adheres to Cal27 cells and markedly boots their proliferation, migratory and invasive capacities under in vitro.
【Key words】 Tongue squamous cell carcinoma; C. leadbetteri; Proliferation; Migration; Invasion

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