Tsutsumi T, Ido A, Nakao K, Hamasaki K, Kato Y, Ohtsuru A, Nakata K, Tamaoki T, Nagataki S
First Department of Internal Medicine, Nagasaki University School of Medicine, Japan.
Gastroenterology. 1994 Aug;107(2):499-504. doi: 10.1016/0016-5085(94)90177-5.
BACKGROUND/AIMS: Butyrate, a product of colonic bacterial flora, functions as an antiproliferative agent and induces cell differentiation in a variety of cell types. In the present study, the effects of butyrate on cell growth and expression of alpha-fetoprotein (AFP) and albumin genes in HuH-7 human hepatoma cells were investigated.
The HuH-7 cells were treated with sodium butyrate (0-1 mmol/L), and numbers of viable cells were counted at 24, 48, and 72 hours after treatment. To elucidate the effects of sodium butyrate on AFP and albumin gene expression, Northern blotting and transient chloramphenicol acetyltransferase plasmid transfection experiments were performed.
Cell growth was dose dependently inhibited by sodium butyrate. By Northern blot analysis, the level of AFP messenger RNA was reduced by treatment with sodium butyrate, whereas the level of albumin messenger RNA was elevated by this treatment. In transient chloramphenicol acetyltransferase plasmid transfection experiments, sodium butyrate repressed the AFP promoter activity but did not change the AFP enhancer or silencer activities. In contrast, the albumin promoter activity was stimulated by sodium butyrate.
These results suggest that butyrate leads to the reciprocal differentiating regulation of AFP and albumin gene expression at the transcriptional level in human hepatoma cells.
背景/目的：丁酸盐是结肠细菌菌群的产物，作为一种抗增殖剂，可诱导多种细胞类型发生细胞分化。在本研究中，研究了丁酸盐对HuH-7人肝癌细胞生长以及甲胎蛋白（AFP）和白蛋白基因表达的影响。
用丁酸钠（0 - 1 mmol/L）处理HuH-7细胞，并在处理后24、48和72小时对活细胞数量进行计数。为阐明丁酸钠对AFP和白蛋白基因表达的影响，进行了Northern印迹和瞬时氯霉素乙酰转移酶质粒转染实验。
丁酸钠剂量依赖性地抑制细胞生长。通过Northern印迹分析，丁酸钠处理降低了AFP信使核糖核酸的水平，而该处理提高了白蛋白信使核糖核酸的水平。在瞬时氯霉素乙酰转移酶质粒转染实验中，丁酸钠抑制了AFP启动子活性，但未改变AFP增强子或沉默子活性。相反，丁酸钠刺激了白蛋白启动子活性。
这些结果表明，丁酸盐在人肝癌细胞中导致AFP和白蛋白基因表达在转录水平上的相互分化调节。
