Formation of insulin-producing cells from pancreatic acinar AR42J cells by hepatocyte growth factor
Formation of insulin-producing cells from pancreatic acinar AR42J cells by hepatocyte growth factor
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DOI：
10.1210/en.137.9.3969
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发表时间：
1996-09-01
期刊：
影响因子：
4.8
通讯作者：
Kojima, I
中科院分区：
文献类型：
--
作者：
Mashima, H;Shibata, H;Kojima, I
胰腺AR 42 J细胞来源于腺泡细胞，表达外分泌和神经内分泌特性。我们最近已经证明，这些细胞转化成胰岛素产生细胞在体外用激活素A和β细胞素处理后。在这里，我们研究了肝细胞生长因子（HGF）在这些细胞中的作用。当AR 42 J细胞与HGF孵育时，DNA合成减弱，淀粉酶含量以浓度依赖性方式降低。HGF处理的细胞延长过程，但使用微管蛋白抗体未观察到束形成。逆转录-PCR分析显示，胰岛素和胰多肽（PP）的信使RNA在经过HGF处理的AR 42 J细胞中表达，但在未经处理的AR 42 J细胞中不表达。免疫细胞化学分析表明，约3%的HGF处理的细胞与抗胰岛素抗体染色，一些也与抗PP抗体染色。当AR 42 J细胞暴露于激活素A和HGF的组合时，细胞延长更长的突起，并且超过10%的突起被抗胰岛素抗体染色。在这些细胞中，表达胰岛素、PP、葡萄糖转运蛋白2和葡萄糖激酶的信使RNA，但不表达胰高血糖素或生长抑素。获得AR 42 J细胞的亚克隆，AR 42 J-B13。大多数AR 42 J-B13细胞在与激活素A和HGF孵育后转化为胰岛素产生细胞。胰岛素分泌增加甲苯磺丁脲，去极化浓度的钾，卡巴胆碱，胰高血糖素样肽-1在这些细胞。这些结果表明，HGF降低了AR 42 J细胞的腺泡细胞样性质，并将其转化为胰岛素产生细胞。激活素A可明显增强HGF的作用。
Pancreatic AR42J cells are derived from acinar cells and express both exocrine and neuroendocrine properties. We have recently shown that these cells convert into insulin-producing cells in vitro after treatment with activin A and betacellulin. Here, we investigated the effect of hepatocyte growth factor (HGF) in those cells. When AR42J cells were incubated with HGF, DNA synthesis was attenuated, and the amylase content was reduced in a concentration-dependent manner. HGF-treated cells extended processes, but bundle formation was not observed using an antibody against tubulin. Reverse transcription-PCR analysis revealed that messenger RNAs for both insulin and pancreatic polypeptide (PP) were expressed in HGF-treated, but not naive, AR42J cells. Immunocytochemical analysis indicated that approximately 3% of the HGF-treated cells were stained with antiinsulin antibody, and some were also stained with anti-PP antibody. When AR42J cells were exposed to a combination of activin A and HGF, cells extended longer processes, and over 10% of them were stained with antiinsulin antibody. In these cells, messenger RNAs for insulin, PP, glucose transporter 2, and glucokinase, but not those for glucagon or somatostatin, were expressed. A subclone of AR42J cells, AR42J-B13, was obtained. Most of the AR42J-B13 cells converted to insulin-producing cells after the incubation with activin A and HGF. Insulin secretion was augmented by tolbutamide, depolarizing concentrations of potassium, carbachol, and glucagon-like peptide-1 in these cells. These results indicate that HGF reduces the acinar cell-like property of AR42J cells and converts them into insulin-producing cells. The effect of HGF was markedly enhanced by activin A.