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大鼠乳糖酶基因启动子在转染的人结肠癌细胞中的活性。

Activity of the rat lactase gene promoter in transfected human colon cancer cells.

作者信息

Boukamel R, Neuville P, Duluc I, Freund J N

机构信息

INSERM U. 381, Strasbourg, France.

出版信息

C R Acad Sci III. 1995 Nov;318(11):1133-40.

PMID:8574790
Abstract

The promoter activity of the upstream region of the rat small intestinal lactase-phlorizin hydrolase gene has been analysed by transfection in the human colon cancer cell line Caco-2. A 0.9 kb mRNA, corresponding to the CAT reporter gene, was synthesized from the transcription start site of the LPH gene. The rate of expression, determined by semi-quantitative RT-PCR, was very low, and depended on the length of the promoter fragment in front of the reporter gene. By immunocytology, we found that the low level of expression resulted from the low number of cells (about 1%) in which CAT was produced. The endogenous lactase was present in 10-20% of the cells in culture, and evidence is provided that most cells that expressed CAT did not co-express the endogenous lactase. We conclude from this study that the rat small intestinal LPH gene promoter is active in the human Caco-2 colon cancer cells. Hence Caco-2 cells constitute an in vitro model to analyse the basic molecular mechanisms involved in the gene transcription of intestinal digestive enzymes. Yet, the mosaic expression of the endogenous lactase and of the reporter gene under the control of the rat LPH gene promoter, suggests that Caco-2 cells may present specific regulatory mechanisms of expression of small intestinal enzymes, possibly in relation to their tumourous origin.

摘要

通过转染人结肠癌细胞系Caco-2,对大鼠小肠乳糖酶-根皮苷水解酶基因上游区域的启动子活性进行了分析。从LPH基因的转录起始位点合成了一个与CAT报告基因相对应的0.9 kb mRNA。通过半定量RT-PCR测定的表达率非常低,并且取决于报告基因前启动子片段的长度。通过免疫细胞化学,我们发现低水平的表达是由于产生CAT的细胞数量较少(约1%)所致。内源性乳糖酶存在于培养细胞的10%-20%中,并且有证据表明大多数表达CAT的细胞不共表达内源性乳糖酶。我们从这项研究中得出结论,大鼠小肠LPH基因启动子在人Caco-2结肠癌细胞中具有活性。因此,Caco-2细胞构成了一个体外模型,用于分析参与肠道消化酶基因转录的基本分子机制。然而,内源性乳糖酶和大鼠LPH基因启动子控制下的报告基因的镶嵌表达表明,Caco-2细胞可能呈现小肠酶表达的特定调控机制,这可能与其肿瘤起源有关。

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