Peptide production and secretion in GLUTag, NCI-H716, and STC-1 cells: a comparison to native L-cells.
Peptide production and secretion in GLUTag, NCI-H716, and STC-1 cells: a comparison to native L-cells.
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DOI：
10.1530/jme-15-0293
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发表时间：
2016-04
影响因子：
3.5
通讯作者：
Holst JJ
中科院分区：
文献类型：
--
作者：
Kuhre RE;Wewer Albrechtsen NJ;Deacon CF;Balk-Møller E;Rehfeld JF;Reimann F;Gribble FM;Holst JJ
GLUTag、NCI-H716和STC-1是广泛用于研究胰高血糖素样肽（GLP-1）分泌机制的细胞系，但它们与天然L细胞相似的程度尚不清楚。我们使用经验证的14种不同激素的免疫测定法分析肽含量（裂解样品; n=9，来自不同的传代次数）或响应缓冲液（基线）的肽分泌，以及用50 mM KCl或10 mM葡萄糖+ 10 μM毛喉素/3-异丁基-1-甲基黄嘌呤刺激后（n=6，也是不同的传代次数）。所有细胞系均以与人肠道所述相似的模式（激素原转化酶（PC）1/3依赖性）产生胰高血糖素原并将其加工成GLP-1、GLP-2、胰高血糖素原和胃泌酸调节素。所有三种细胞系均显示GLP-1和GLP-2的基础分泌，其在刺激后增加。与新鲜分离的鼠L细胞相反，所有细胞系也表达PC 2并分泌大量胰高血糖素。神经降压素和生长抑素的存储量低，分泌并不一致的刺激增加。STC-1细胞分泌的葡萄糖依赖性促胰岛素多肽（GIP）明显高于GLP-1细胞（P<0.01），KCl使其分泌增加（P<0.05）。在任何细胞系中均未检测到通常与GLP-1共定位于远端L细胞中的PYY。GLUTag和STC-1细胞也表达血管活性肠肽，但没有表达胰腺多肽或胰岛素。GLUTag含有并分泌大量胆囊收缩素，而NCI-H716不储存该肽，STC-1含有少量。我们的研究结果表明，L-细胞的细胞系模型中的激素生产与天然L-细胞的相似性有限。
GLUTag, NCI-H716 and STC-1 are cell lines that are widely used to study mechanisms underlying secretion of glucagon-like peptide (GLP-1), but the extent to which they resemble native L-cells is unknown. We used validated immunoassays for 14 different hormones to analyze peptide content (lysis samples; n=9 from different passage numbers) or peptide secretion in response to buffer (baseline), and after stimulation with 50 mM KCl or 10 mM glucose + 10 μM forskolin/3-isobutyl-1-methylxanthine (n=6 also different passage numbers). All cell lines produced and processed proglucagon into GLP-1, GLP-2, glicentin and oxyntomodulin in a pattern (prohormone convertase (PC)1/3 dependent) similar to that described for human gut. All three cell lines showed basal secretion of GLP-1 and -2 which increased after stimulation. In contrast to freshly isolated murine L-cells, all lines also expressed PC2 and secreted large amounts of pancreatic glucagon. Neurotensin and somatostatin storage was low and secretion was not consistently increased by stimulation. STC-1 cells released more glucose-dependent insulinotropic polypeptide (GIP) than GLP-1 at baseline (P<0.01) and KCl elevated its secretion (P<0.05). PYY, which normally co-localizes with GLP-1 in distal L-cells, was not detected in any of the cell lines. GLUTag and STC-1 cells also expressed vasoactive intestinal peptide, but none expressed pancreatic polypeptide or insulin. GLUTag contained and secreted large amounts of cholecystokinin while NCI-H716 did not store this peptide and STC-1 contained low amounts. Our results show that hormone production in cell line models of the L-cell have limited similarity to the natural L-cells.