Zhou C Y, Li X M, Shan S, Jia L F, Huang Z L
Department of Otolaryngology,Chinese People's Liberation Army 401 Hospital,Qingdao,266071,China.
Department of Otolaryngology Head and Neck Surgery, Bethune International Peace Hospital.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2017 Apr 5;31(7):524-528. doi: 10.13201/j.issn.1001-1781.2017.07.008.
To investigate metformin's effect on chemosensitivity of chemotherapeutic drug 5-fluorouracil in laryngocarcinoma Hep-2 cells. Investigate the variation trend of protein expression of AMPK pathway in the combined effect.Laryngocarcinoma Hep-2 cells were treated with different concentrations of 5-fluorouracil in vitro together with or without metformin for 72 h. Use MTT assay to investigate the influence on the inhibition rate to Hep-2 cells. Hep-2 cells were treated with cisplatin, 5-fluorouracil or paclitaxel with or without metformin. Use Western blot assay to investigate the expression level of AMPKα, P21 or Cyclin D1 protein. 5-fluorouracil and metformin could inhibit the proliferation of Hep-2 cells. 5-fluorouracil in low concentration combined with metformin could increase the proliferation inhibition rate of Hep-2 cells. In the circumstances of using 5-fluorouracil in high concentration with metformin , the cell proliferation inhibition rate of combining group makes no differences with the single-drug group. The combination of metformin and 5-fluorouracil produced an antagonism action in Hep-2 cells.Western blot assay showed that metformin, cisplatin, 5-fluorouracil could have caused the increase of expression level of AMPK-α, P21 and Cyclin D1 in Hep-2 cells while Paclitaxel could have cause the decrease of expression level of Cyclin D1. Using combined drug could cause the change of protein expression. 5-fluorouracil has been found to inhibit the proliferation of Hep-2 cells. Metformin has an antagonism on the anticancer effect to 5-fluorouracil in Hep-2 cells, and this antagonistic effect occurred partially through molecular signal pathways of AMPK-α, P21 and Cyclin D1 and it's significantly related to the cell cycle arrest.
探讨二甲双胍对喉癌Hep-2细胞化疗药物5-氟尿嘧啶化疗敏感性的影响。研究联合作用时AMPK通路蛋白表达的变化趋势。体外将喉癌Hep-2细胞用不同浓度的5-氟尿嘧啶处理72小时，同时加入或不加入二甲双胍。采用MTT法检测对Hep-2细胞抑制率的影响。将Hep-2细胞用顺铂、5-氟尿嘧啶或紫杉醇处理，同时加入或不加入二甲双胍。采用蛋白质印迹法检测AMPKα、P21或细胞周期蛋白D1蛋白的表达水平。5-氟尿嘧啶和二甲双胍可抑制Hep-2细胞的增殖。低浓度5-氟尿嘧啶联合二甲双胍可提高Hep-2细胞的增殖抑制率。高浓度5-氟尿嘧啶与二甲双胍联合使用时，联合组细胞增殖抑制率与单药组无差异。二甲双胍与5-氟尿嘧啶联合对Hep-2细胞产生拮抗作用。蛋白质印迹法显示，二甲双胍、顺铂、5-氟尿嘧啶可使Hep-2细胞中AMPK-α、P21和细胞周期蛋白D1表达水平升高，而紫杉醇可使细胞周期蛋白D1表达水平降低。联合用药可引起蛋白表达的改变。已发现5-氟尿嘧啶可抑制Hep-2细胞的增殖。二甲双胍对5-氟尿嘧啶在Hep-2细胞中的抗癌作用具有拮抗作用，且这种拮抗作用部分通过AMPK-α、P21和细胞周期蛋白D1的分子信号通路发生，且与细胞周期阻滞显著相关。
