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肿瘤坏死因子-α通过Wnt/β-catenin信号通路对胆囊癌细胞增殖的作用

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

    摘要
  • 摘要:
    目的 探讨肿瘤坏死因子-α(TNF-α)通过Wnt/β-catenin信号通路对胆囊癌细胞系NOZ的增殖作用, 并探讨其相关分子机制。
    方法 采用CCK-8实验和生长曲线实验测得TNF-α对胆囊癌细胞系NOZ增殖作用的最佳浓度和时间; 采用流式细胞术检测TNF-α对胆囊癌细胞系NOZ周期的影响; 应用Western blot方法检测胆囊癌细胞系NOZ中Wnt/β-catenin信号通路相关基因(β-catenin、c-myc、cyclinD1)蛋白的表达; 采用平板克隆形成实验测定胆囊癌细胞系NOZ的增殖能力。
    结果 CCK-8实验法和生长曲线实验测得TNF-α对胆囊癌细胞系NOZ增殖作用的最佳浓度是20 ng/mL, 最佳时间是48 h; 在TNF-α20 ng/mL作用下, G1期细胞数目减少, S期细胞占比增加, G2期细胞占比减少; TNF-α可促进胆囊癌细胞系NOZ中Wnt/β-catenin信号通路相关基因(β-catenin、c-myc、cyclinD1)蛋白的表达; 经TNF-α(20 ng/mL)处理的胆囊癌细胞系NOZ克隆数量大于对照组，差异有统计学意义(P<0.05)。
    结论 TNF-α可能通过Wnt/β-catenin信号通路促进胆囊癌细胞系NOZ的增殖，促进胆囊癌发展。

     

    Abstract:
    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.

     

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