Pdx-1 or Pdx-1-VP16 protein transduction induces beta-cell gene expression in liver-stem WB cells.
Pdx-1 or Pdx-1-VP16 protein transduction induces beta-cell gene expression in liver-stem WB cells.
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DOI：
10.1186/1756-0500-2-3
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发表时间：
2009-01-09
影响因子：
1.8
通讯作者：
Bösch S
中科院分区：
文献类型：
--
作者：
Delisle JC;Martignat L;Dubreil L;Saï P;Bach JM;Louzier V;Bösch S
胰腺十二指肠同源盒-1 (Pdx-1) 或 Pdx-1-VP16 基因转移已被证明可诱导体外大鼠肝干 WB 细胞转化为胰腺内分泌前体细胞。高葡萄糖条件是进一步分化为功能性胰岛素产生细胞所必需的。由于固有的蛋白质转导结构域 (PTD)，Pdx-1 能够渗透不同的细胞类型。在这项研究中，我们评估了 Pdx-1 或 Pdx-1-VP16 蛋白转导后 WB 细胞的肝脏到胰腺的转化。 WB 细胞在含有 Pdx-1 或 Pdx-1-VP16 重组蛋白的高葡萄糖培养基中生长两周。通过胰腺内分泌基因的 RT-PCR 分析 β 样细胞定型。我们发现高糖培养中的WB细胞自发表达胰腺内分泌基因（Pdx-1、Ngn3、Nkx2.2、Kir6.2）。仅在 Pdx-1(-VP16) 蛋白存在的情况下，它们才能进一步分化为表达与内分泌胰腺发育（Ngn3、NeuroD、Pax4、Nkx2.2、Nkx6.1、Pdx-1）和 β 细胞功能（Glut-2、Kir6.2、胰岛素）相关基因的 β 样细胞。这些结果表明，Pdx-1(-VP16) 蛋白转导有助于体外肝脏到胰腺的转化，并且是糖尿病细胞治疗的 β 细胞工程基因治疗的替代方案。
Pancreatic duodenal homeobox-1 (Pdx-1) or Pdx-1-VP16 gene transfer has been shown to induce in vitro rat liver-stem WB cell conversion into pancreatic endocrine precursor cells. High glucose conditions were necessary for further differentiation into functional insulin-producing cells. Pdx-1 has the ability to permeate different cell types due to an inherent protein transduction domain (PTD). In this study, we evaluated liver-to-pancreas conversion of WB cells following Pdx-1 or Pdx-1-VP16 protein transduction. WB cells were grown in high glucose medium containing Pdx-1 or Pdx-1-VP16 recombinant proteins for two weeks. β-like cell commitment was analysed by RT-PCR of pancreatic endocrine genes. We found that WB cells in high glucose culture spontaneously express pancreatic endocrine genes (Pdx-1, Ngn3, Nkx2.2, Kir6.2). Their further differentiation into β-like cells expressing genes related to endocrine pancreas development (Ngn3, NeuroD, Pax4, Nkx2.2, Nkx6.1, Pdx-1) and β-cell function (Glut-2, Kir6.2, insulin) was achieved only in the presence of Pdx-1(-VP16) protein. These results demonstrate that Pdx-1(-VP16) protein transduction is instrumental for in vitro liver-to-pancreas conversion and is an alternative to gene therapy for β-cell engineering for diabetes cell therapy.