Role of tumor necrosis factor-α in promoting gallbladder cancer cell proliferation through Wnt/β-catenin signaling pathway

Objective To investigate the proliferation effect of tumor necrosis factor-α(TNF-α) on gallbladder cancer cell line NOZ through Wnt/β-catenin signaling pathway, and to explore its molecular mechanism.

Methods CCK-8 experiment and growth curve experiment were used to determine the optimal concentration and time of TNF-α on the proliferation of gallbladder cancer cell line NOZ. The effect of TNF-α on cycle of gallbladder cancer cell line NOZ was detected by flow cytometry. Western blot was used to detect the expression of Wnt/β-catenin signaling pathway related genes (β-catenin, c-myc, cyclinD1) in gallbladder cancer cell line NOZ. Proliferation ability of gallbladder cancer cell line NOZ was determined by plate clone formation experiment.

Results CCK-8 experiment and growth curve experiment showed that the optimal concentration and time of TNF-α on the proliferation of gallbladder cancer cell line NOZ were 20 ng/mL and 48 h; TNF-α decreased the number of cells in G₁ phase, increased the proportion of cells in S phase and decreased the proportion of cells in G₂ phase under the action of 20 ng/mL (P<0.05). TNF-α can promote the expression of Wnt/β-catenin signaling pathway related genes (β-catenin, c-myc, cyclinD1) in gallbladder cancer cell line NOZ (P<0.05). The number of NOZ clones in gallbladder cancer cell line treated with TNF-α(20 ng/mL) was higher than that in control group (P<0.05).

Conclusion TNF-α may promote the proliferation of gallbladder cancer cell line NOZ through Wnt/β-catenin signaling pathway.