Dudas Jozsef, Elmaouhoub Abderrahim, Mansuroglu Tümen, Batusic Danko, Tron Kyrylo, Saile Bernhard, Papoutsi Maria, Pieler Tomas, Wilting Joerg, Ramadori Giuliano
Department of Internal Medicine, Section of Gastroenterology and Endocrinology, Georg-August-University Göttingen, Robert-Koch-Strasse 40, 37075 Goettingen, Germany.
Histochem Cell Biol. 2006 Nov;126(5):549-62. doi: 10.1007/s00418-006-0191-4. Epub 2006 Jun 13.
The aim of this study was to analyse the changes of Prospero-related homeobox 1 (Prox1) gene expression in rat liver under different experimental conditions of liver injury, regeneration and acute phase reaction, and to correlate it with that of markers for hepatoblasts, hepatocytes, cholangiocytes and oval cells. Gene expression was studied at RNA level by RT-PCR, and at protein level by immunohistochemistry. At embryonal stage of rat liver development (embryonal days (ED) 14-16) hepatoblasts were found to be Prox1(+)/Cytokeratin (CK) 19(+) and alpha-fetoprotein (AFP)(+), at this stage Prox1(-)/CK19(+)/AFP(-) small cells (early cholangiocytes?) were identified. In fetal liver (ED 18-22) hepatoblasts were Prox1(+)/CK19(-)/AFP(+). CK7(+) cholangiocytes were detected at this stage, and they were Prox1(-)/AFP(-). In the adult liver hepatocytes were Prox1(+)/CK19(-)/CK7(-)/AFP(-), cholangiocytes were CK19(+) and/or CK7(+) and AFP(-)/Prox1(-). In models of liver damage and regeneration Prox1 remained a stable marker of hepatocytes. After 2-acetyl-aminofluorene treatment with partial hepatectomy (AAF/PH) the amount of Prox1 specific transcripts was low in the liver, when CK19 and AFP gene expression was high, and at no time point AFP(+)/CK19(+ )"oval cells" were found to be Prox1(+). However, a few Prox1(+)/CK19(+) and a few Prox1(+)/CK7(+ )cells were identified in the liver of AAF/PH-animals, which may represent precursors of hepatocytes, or a precancerous state.
本研究的目的是分析在肝损伤、再生和急性期反应的不同实验条件下,大鼠肝脏中Prospero相关同源盒1(Prox1)基因表达的变化,并将其与肝母细胞、肝细胞、胆管细胞和卵圆细胞的标志物表达相关联。通过逆转录聚合酶链反应(RT-PCR)在RNA水平研究基因表达,通过免疫组织化学在蛋白质水平研究基因表达。在大鼠肝脏发育的胚胎阶段(胚胎日(ED)14 - 16),发现肝母细胞为Prox1(+)/细胞角蛋白(CK)19(+)和甲胎蛋白(AFP)(+),在此阶段鉴定出Prox1(-)/CK19(+)/AFP(-)的小细胞(早期胆管细胞?)。在胎儿肝脏(ED 18 - 22)中,肝母细胞为Prox1(+)/CK19(-)/AFP(+)。在此阶段检测到CK7(+)胆管细胞,它们为Prox1(-)/AFP(-)。在成年肝脏中,肝细胞为Prox1(+)/CK19(-)/CK7(-)/AFP(-),胆管细胞为CK19(+)和/或CK7(+)且AFP(-)/Prox1(-)。在肝损伤和再生模型中,Prox1仍然是肝细胞的稳定标志物。在2 - 乙酰氨基芴处理并部分肝切除(AAF/PH)后,肝脏中Prox1特异性转录本的量较低,此时CK19和AFP基因表达较高,且在任何时间点均未发现AFP(+)/CK19(+)“卵圆细胞”为Prox1(+)。然而,在AAF/PH动物的肝脏中鉴定出少数Prox1(+)/CK19(+)和少数Prox1(+)/CK7(+)细胞,它们可能代表肝细胞的前体,或癌前状态。