Rodolosse A, Chantret I, Lacasa M, Chevalier G, Zweibaum A, Swallow D, Rousset M
INSERM U178 and Université Paris-Sud, Villejuif, France.
Biochem J. 1996 Apr 1;315 ( Pt 1)(Pt 1):301-6. doi: 10.1042/bj3150301.
We have previously shown that glucose can exert a repressive effect on the transcription of the sucrase-isomaltase (SI) gene in the differentiated enterocyte-like human colon carcinoma cell lines HT-29 and Caco-2. To characterize the region through which glucose exerts this effect, three different-length fragments of the 5'-flanking region of the human SI gene were linked to the reporter gene luciferase in an episomal vector carrying a hygromycin resistance gene. These fragments were used for transfection into a clone of the Caco-2 cell line, PF11, which has high glucose consumption and only expresses SI at high levels when cultured in the presence of a low supply of glucose. By using the stably transformed PF11 cells grown either in standard high glucose (25 mM) or in low glucose (1 mM) it was possible to show that the smallest fragment of the SI promoter, extending from bases -370 to +30, contains all the information required for the glucose repression of the reporter gene luciferase.
我们之前已经表明,葡萄糖可对分化的肠上皮样人结肠癌细胞系HT-29和Caco-2中的蔗糖酶-异麦芽糖酶(SI)基因转录产生抑制作用。为了确定葡萄糖发挥此效应的区域,将人SI基因5'-侧翼区的三个不同长度片段与携带潮霉素抗性基因的游离载体中的报告基因荧光素酶相连。这些片段用于转染Caco-2细胞系的一个克隆PF11,该克隆具有高葡萄糖消耗,并且仅在低葡萄糖供应条件下培养时才高水平表达SI。通过使用在标准高葡萄糖(25 mM)或低葡萄糖(1 mM)中生长的稳定转化的PF11细胞,有可能表明SI启动子的最小片段,从碱基-370延伸至+30,包含报告基因荧光素酶葡萄糖抑制所需的所有信息。