[Biological effect of nicotine on Cal27 cell line in patients with tongue squamous cell carcinoma].

PURPOSE: To investigate the effect of nicotine on Cal27 cell line in patients with tongue squamous cell carcinoma.

METHODS

Tongue squamous cell carcinoma Cal27 cells were subcultured, and the logarithmic growth phase cells were selected and divided into blank control group, nicotine-treated group and α7 nicotinic acetylcholine receptor inhibition-treated group (α7nAChR inhibition group). Cells in the blank control group received no treatment; cells in nicotine group received nicotine treatment, and cells in α7nAChR inhibition group were treated nicotine combined with α-bungarotoxin (α-BTX). The treatments lasted for 1 week. Cell morphology of blank control group and nicotine group was observed under electron microscope, cell proliferation was detected by CCK-8 kit, cell migration ability was detected by scratch test, cell invasion ability of blank control group and nicotine group was detected by Transwell chamber. The relative expression levels of Wnt signaling pathway proteins in nicotine group, blank control group and α7nAChR inhibition group were determined by Western blot. The data were statistically analyzed using SPSS 20.0 software package.

RESULTS

Microscope showed that the rapidly adherent cells were uniformed polygon shape with large nucleus, and there was no significant difference among 3 groups. After 96 hours of incubation, the number of cells in nicotine group was significantly higher than that in the blank control group and α7nAChR inhibition group (P<0.05). The degree of scratch healing in nicotine group was significantly higher than that in the blank control group and α7nAChR inhibition group (P<0.05). The number of cells passing through the chamber in nicotine group was significantly higher than that of the blank control group andα7nAChR inhibition group (P<0.05). The relative expression levels of Wnt pathway proteins including β-catenin, c-Myc, p-GSK3β and Ror2 were significantly higher in nicotine group than those in the blank control group and α7nAChR inhibition group (P<0.05).

CONCLUSIONS

Nicotine can enhance proliferation, migration and invasion of tongue squamous cell carcinoma Cal27 cells by activating Wnt signaling pathway.