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目的:检测5-氟尿嘧啶联合二甲双胍对喉癌Hep-2细胞增殖抑制作用及化疗敏感影响。探究二甲双胍联合顺铂、紫杉醇、5-氟尿嘧啶作用于Hep-2细胞AMPK通路信号分子的表达变化。方法:体外培养Hep-2细胞,二甲双胍联合5-氟尿嘧啶作用细胞72h,MTT检测增殖情况。二甲双胍联合顺铂、5-氟尿嘧啶、紫杉醇作用细胞72h,Western blot检测P21、Cyclin D1、AMPK-α表达。结果:5-氟尿嘧啶以及二甲双胍可抑制Hep-2细胞增殖。二甲双胍联合低浓度5-氟尿嘧啶,细胞生长抑制率升高;二甲双胍联合高浓度5-氟尿嘧啶生长抑制率无明显差异,两药在Hep-2细胞中相互拮抗。Western blot显示,顺铂、5-氟尿嘧啶增加细胞中上述蛋白表达,紫杉醇减少细胞中Cyclin D1表达。对比单独用药组,联用二甲双胍均引起Hep-2细胞中上述蛋白表达变化。结论:5-氟尿嘧啶抑制Hep-2细胞增殖,二甲双胍拮抗5-氟尿嘧啶对Hep-2细胞抗肿瘤效应,其拮抗作用及对顺铂、紫杉醇的影响部分通过AMPK、P21及Cyclin D1相关通路发生作用且与周期阻滞存在密切关系。
Objective: To investigate the effects of 5-fluorouracil combined with metformin on the proliferation and chemosensitivity of Hep-2 laryngeal carcinoma cells. To investigate the effect of metformin combined with cisplatin, paclitaxel and 5-fluorouracil on the expression of AMPK signaling in Hep-2 cells. Methods: Hep-2 cells cultured in vitro, metformin combined with 5-fluorouracil-treated cells 72h, MTT assay proliferation. Metformin combined with cisplatin, 5-fluorouracil and paclitaxel for 72 hours. Western blot was used to detect the expression of P21, Cyclin D1 and AMPK-α. Results: 5-Fluorouracil and metformin inhibited Hep-2 cell proliferation. Metformin combined with low concentration of 5-fluorouracil, cell growth inhibition rate increased; metformin combined with high concentration of 5-fluorouracil growth inhibition rate was no significant difference between the two drugs in Hep-2 cells antagonistic. Western blot showed that cisplatin and 5-fluorouracil increased the expression of these proteins in cells, and paclitaxel reduced the expression of Cyclin D1 in cells. Compared with the single treatment group, the combination of metformin caused Hep-2 cells in the above protein expression changes. CONCLUSION: 5-Fluorouracil inhibits the proliferation of Hep-2 cells. Metformin antagonizes the antitumor effect of 5-Fluorouracil on Hep-2 cells. Its antagonism and its effects on cisplatin and paclitaxel partly through AMPK, P21 and Cyclin D1 related pathways There is a close relationship with cycle arrest.