In vitro transformation of adult rat hepatic progenitor cells into pancreatic endocrine hormone-producing cells
In vitro transformation of adult rat hepatic progenitor cells into pancreatic endocrine hormone-producing cells
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DOI：
10.1007/s00534-007-1252-3
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发表时间：
2008-05-01
期刊：
影响因子：
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通讯作者：
Hirata, Koichi
中科院分区：
文献类型：
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作者：
Kawasaki, Hiroyuki;Mizuguchi, Toru;Hirata, Koichi
背景/目的。胰腺和十二指肠同源盒因子1（Pdx-1）在启动胰腺内分泌细胞向胰腺内分泌细胞的分化中起重要作用。肝细胞转分化或转化为胰腺内分泌细胞是可行的，因为它们具有相似的细胞起源。本研究的目的是观察小肝细胞（SH）是否能通过外源性Pdx-1基因表达产生胰腺内分泌细胞。培养10 d后，腺病毒（Adt）介导的Pdx-1基因转染SHs。我们进行了蛋白质印迹分析胰腺转录因子的细胞核和逆转录聚合酶链反应（RT-PCR）的胰腺内分泌激素的基因表达。采用共聚焦激光显微扫描分析技术观察SH的转化情况。Adt-Pdx-1基因转染后，胰腺转录因子如Pdx-1、Ngn 3、NeuroD、Nkx 2.2和Pax 6被诱导。基因转染后诱导胰腺内分泌激素（胰岛素、胰高血糖素和生长抑素）的mRNA表达。Pdx-1在细胞核中表达，其中细胞对一种或多种激素和细胞角蛋白（CK）8呈阳性。一些细胞对多种激素呈阳性。葡萄糖负荷试验后，胰岛素水平升高，胰高血糖素水平降低，这与葡萄糖浓度有关。转染Pdx-1基因后，SH转化为具有功能的胰腺内分泌细胞。
Background/Purpose. Pancreatic and duodenal homeobox factor 1 (Pdx-1) plays an important role in initiating differentiation toward pancreatic endocrine cells. The transdifferentiation or transformation of hepatocytes into pancreatic endocrine cells could be feasible, due to their similar cellular origins. Our goal in this study was to see if small hepatocytes (SHs) could give rise to pancreatic endocrine cells via exogenous Pdx-1 gene expression.Methods. SHs were cultured for 10 days before adenovirus (Adt)-mediated Pdx-1 gene transfection. We performed western blot analysis for pancreatic transcription factors in the nuclei and reverse-transcription polymerase chain reaction (RT-PCR) for the gene expression of pancreatic endocrine hormones. Confocal laser microscanning analysis was used to observe the transformation of SHs.Results. Pancreatic transcription factors such as Pdx-1, Ngn3, NeuroD, Nkx2.2, and Pax6 were induced after Adt-Pdx-1 gene transfection. The mRNA expression of pancreatic endocrine hormones (insulin, glucagon, and somatostatin) was induced after the gene transfection. Pdx-1 was expressed in the nucleus, where the cells were positive for one or more of the hormones and cytokeratin (CK) 8. Some cells were positive for multiple hormones. The insulin level increased while the glucagon level decreased after the glucose loading test, depending on the glucose concentration.Conclusions. SHs are transformed into functional pancreatic endocrine cells after Pdx-1 gene transfection.