肿瘤细胞^3H-葡萄糖代谢测量方法的建立

目的检测肿瘤细胞在单位时间内的葡萄糖利用率。方法将数量为2.5×10^5的HCT116结肠癌细胞培养至贴壁,在培养基中加入^3H定位标记（5位碳原子）的葡萄糖,30 min后终止反应,采用阴离子交换树脂滤去大分子代谢产物,通过液体闪烁计数法测量流出液中氚水的每分放射性计数,计算得到单位数目的 HCT116细胞在30min内的葡萄糖代谢率。结果当层析柱内的层析液体积大于5.0 ml时,层析柱对^3H-葡萄糖标准液内葡萄糖的吸附率几乎达到100%（99%）;30 min内3.7×10^5的HCT116细胞的葡萄糖代谢率约为1.23%。结论该方法通过在细胞培养液中加入同位素^3H定位标记的葡萄糖,可以准确定量测定肿瘤细胞的葡萄糖代谢率,从而为研究不同种类肿瘤细胞糖代谢及能量供给提供可靠的实验方法。

Development of the Method of Determining the ^3H-Glucose Metabolism by Tumor Cells

Objective To measure the glucose metabolic rate（GMR） in tumor cells.Methods HCT116 cells were seeded in a density of 2×10^5 and grown to 90%confluence.Then 5- H-glucose was added into the medium.Thirty minutes later,cells were killed while ^3H-H2O was separated from the labeled glucose by column chromatography.Radioactivity of ^3H-H2O was counted using scintillation counter. At last glucose metabolic rate（GMR） in 30 minutes was calculated.Results The chromatography column was found to exclude more than 99%of the total H-glucose from the perfusate if there was more than 5.0 ml of suspension loaded into the column.The glucose metabolic rate（GMR） of 3.7×10^5 HCT116 cells was 1.23%in 30 minutes.Conclusion It is proved that by using this method one can determine the glucose metabolic rate precisely via adding 5- H-glucose into the medium.Using this method, the tumor cells＇ glucose metabolism and energy providing will be studied better.