The promoter of the long variant of collagen XVIII, the precursor of endostatin, contains liver-specific regulatory elements

被引:23
作者
Liétard, J
Théret, N
Rehn, M
Musso, O
Dargère, D
Pihlajaniemi, T
Clément, B
机构
[1] Univ Rennes 1, INSERM U456, Detoxicat & Tissue Repair Unit, F-35043 Rennes, France
[2] Univ Oulu, Bioctr, Collagen Res Unit, Oulu, Finland
[3] Univ Oulu, Dept Med Biochem, Oulu, Finland
关键词
D O I
10.1053/jhep.2000.20066
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
The endostatin precursor collagen XVIII is expressed at high levels in human livers, the main source being hepatocytes. We have studied the regulatory elements in the promoter 2 of the Coll8a1 gene that directs the transcription of the NCl-517 variant of collagen alpha1(XVIII), which is the main form expressed in the liver. The 5'-flanking region of Col18a1 gene was cloned, and a series of 5'-deletions from -3286 bp to +285 bp were linked to the luciferase reporter gene. Transfection experiments in HepG2 cells allowed to identify a silencer-like element containing putative HNF1 and HNF3 sites and activator elements containing stretches of GC-rich sequences. Another putative HNF3 site in close apposition to a NF1/CTF site was localized upstream of the silencer-like element. Cotransfection experiments showed that the Col18a1 promoter 2 was transactivated by Spl and HNF3 alpha. Gel-shift analyses showed that HNF3, NF1/CTF, and Spl-like sites specifically recognized nuclear factors. Super-shift experiments indicated that HNF3 beta was the major form of HNF3 interacting with the HNF3/NF1 site. The well-differentiated hepatoma cell line mhATFS315 transfected with a truncated form of HNF3P, which competitively blocks HNF3 transactivating activity, expressed the Col18a1gene at a very low level. Taken together, these data strongly suggest that Col18a1 is a liver-specific gene. Furthermore, gel-shift analyses performed with nuclear factors prepared from well-differentiated hepatocellular carcinomas showed increased HNF3/NF1 binding activity compared with normal livers. Consequently, the precursor of endostatin might be differently expressed according to the differentiated and/or transformed state of hepatocytes.
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页码:1377 / 1385
页数:9
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