Reengineering salivary gland cells to enhance protein secretion for use in developing artificial salivary gland device
Reengineering salivary gland cells to enhance protein secretion for use in developing artificial salivary gland device
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DOI：
10.1089/ten.2006.0300
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发表时间：
2007-05-01
影响因子：
--
通讯作者：
Palmon, Aaron
中科院分区：
文献类型：
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作者：
Aframian, Doron J.;Amit, Doron;Palmon, Aaron
唾液腺（SG）被认为是外分泌腺，其主要将水分泌到口腔中。然而，它们也表现出较小的内分泌途径流向血流。人工SG装置的概念外分泌液分泌到口腔区域的口干患者先前已经研究。目前研究的目的是检查这种装置用于增强生物活性蛋白分泌的潜力。我们设计了一个质粒，该质粒编码SG特异性信号肽序列，该信号肽序列与通常不分泌的编码报告基因相邻，从而产生嵌合蛋白，并检查了该构建体是否可以增强唾液上皮细胞的分泌。合成N-末端编码表皮生长因子（EGF）序列，并将其插入萤火虫荧光素酶基因的pGL 3对照载体50中，产生pGL 3- EGF信号肽（pGL 3-EGFSP）融合载体。将该载体与含有海肾荧光素酶基因的pRL-CMV载体共转染293细胞（用作对照）和人下颌下腺导管上皮（HSG）、大鼠下颌下腺腺泡上皮（SMIE）和大鼠下颌下腺导管上皮（A5）唾液细胞系。转染的293、SMIE和HSG细胞分别显示出高8倍、18倍和40倍的荧光素酶活性。这些观察结果导致了设想的分泌装置的概念，其可以作为生物活性蛋白的潜在生物泵。
Salivary glands (SGs) are considered exocrine glands, which mainly secrete water into the oral cavity. Nevertheless, they also exhibit a smaller endocrine secretory pathway toward the bloodstream. The concept of an artificial SG device for exocrine fluid secretion into the oral region in xerostomic patients has been previously studied. The purpose of the current study was to examine the potential of such a device for enhancing bioactive protein secretion. We engineered a plasmid encoding a SG-specific signal peptide sequence adjacent to a normally nonsecreted encoded reporter gene creating a chimera protein, and examined if this construct can enhance secretion from salivary epithelial cells. An N-terminal encoding epidermal growth factor (EGF) sequence was synthesized and inserted into a pGL3 control vector 50 of a firefly luciferase gene, creating a pGL3- EGF signal peptide (pGL3-EGFSP) fused vector. This vector was cotransfected with a pRL-CMV vector containing a Renilla luciferase gene, in 293 cells (serving as controls), and human submandibular gland ductal epithelial (HSG), rat submandibular gland acinar epithelial (SMIE), and rat submandibular gland ductal epithelial (A5) salivary cell lines. The transfected 293, SMIE, and HSG cells showed 8-, 18-, and 40-fold higher luciferase activity, respectively. These observations lead to the concept of an envisioned secretory device, which can serve as a potential biological pump for bioactive proteins.