二甲双胍激活AMPK、调控细胞周期抑制裸鼠人食管癌移植瘤生长-消化内科专业论文.docxVIP

Abstract Background and Objectives： The biguanide antihyperglycemic drug metformin is the most widely used for the treatment of type 2 diabetes mellitus. In recent years, the researches have shown that metformin not only reduces the blood glucose levels, but also decrease morbidity and mortality of cancer. And that metformin induces cell cycle arrest by activating AMP-activated protein kinase (AMPK) is demonstrated to be the key mechanisms responsible for its anti-tumor action. But there is rare report about whether or how metformin inhibits growth of esophageal carcinoma xenografts and affects the cell cycle. Our previous in vitro work indicated that metformin induced cell cycle arrest in G0/G1 phase to inhibit proliferation of esophageal carcinoma cell lines. In this study, we investigated the effects on growth inhibition and cell cycle regulation of metformin in esophageal carcinoma xenografts in nude mice and its molecule mechanism. This will offer a new theoretical basis for prevention and treatment of esophageal carcinoma in clinical. Materials and methods： A human esophageal carcinoma cell line, EC109 was cultured in RPMI-1640 with 10% fetal bovine serum supplemented with 100U/ml penicillin and 100U/ml streptomycin. EC109 cells were harvested, and 1×106 cells were implanted subcutaneously into the left and the right flanks of each eight-week-old male athymic nude mice. Two weeks after implantation, animal were randomized into control (normal saline was given daily by gavage) and treatment groups (metformin was given daily by gavage at 250 mg/kg body weight). 5 weeks later, all the animals were sacrificed and the tumors were collected. Western Blot was performed to detect the phosphorylation of AMPK and expression of cell cycle regulatory proteins. Immunohistochemistry was used to detect p53 and p21CIP1 expression on the tissue sections. Before and after intragastric administration，the body weight and the blood glucose of nude mice were detected.