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目的 研究丁酸钠对永生化食管上皮的增殖、分化和凋亡的作用。方法 用HPV18E6E7诱发的人胚食管上皮永生化细胞株SHEE ,培养在 5 0ml培养瓶和 2 4孔培养板 ,实验组分别加入1mmol/L和 5mmol/L丁酸钠 ,对照组未加药 ,作用 4d。统计细胞克隆数 ,细胞超微结构用电镜检查 ;细胞周期和凋亡细胞数用流式细胞仪检查 ;Ki 6 7、细胞角蛋白用免疫组织化学SP法检查 ;激光共聚焦扫描显微镜检查用鬼臼毒素标记的F 肌动蛋白。结果 加入 1mmol/L和 5mmol/L丁酸钠 4d克隆形成率分别为 6 5 5 %和 2 5 5 % ,比对照组 73 5 %减少。细胞周期检查 1mmol/L组S期细胞明显减少 (4 6 % ) ,多停留在G0 /G1期 (83 8% )。与对照组比较 ,1mmol/L组细胞Ki 6 7表达降低 ,F 肌动蛋白和角蛋白表达增加 ,5mmol/L组细胞凋亡明显增多。结论 丁酸钠可以诱导SHEE细胞增殖停滞和细胞凋亡 ,并有促细胞分化作用。其与用药剂量和时间有关。
Objective To study the effects of sodium butyrate on the proliferation, differentiation and apoptosis of immortalized esophageal epithelium. Methods Human embryonic esophageal epithelial immortalized cell line SHEE induced by HPV18E6E7 was cultured in 50 ml culture flasks and 24 well culture plates. The experimental groups were added with 1 mmol/L and 5 mmol/L sodium butyrate, respectively. The control group did not add drugs. 4d. The number of cell clones was counted. The ultrastructure of the cells was examined by electron microscopy; the cell cycle and number of apoptotic cells were examined by flow cytometry; Ki 6.7, cytokeratin was examined by SP immunohistochemistry; confocal laser scanning microscopy was used to examine ghosts. Scorpion toxin labeled F-actin. Results The colony formation rates of 1 mmol/L and 5 mmol/L sodium butyrate for 4 days were 6.55% and 25.5%, respectively, which was 73.5% lower than that of the control group. In the cell cycle test, 1 mmol/L group S-phase cells were significantly reduced (46%), and most of them remained in G0/G1 phase (83%). Compared with the control group, the expression of Ki 67 in 1 mmol/L group was decreased, the expression of F-actin and keratin was increased, and the apoptosis in 5 mmol/L group was significantly increased. Conclusion Sodium butyrate can induce the proliferation arrest and apoptosis of SHEE cells and promote cell differentiation. It is related to the amount of medication and time.