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首页> 外文期刊>Cellular & molecular biology letters. >Rab3D regulates amylase levels, not agonist-induced amylase release, in AR42J cells.
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Rab3D regulates amylase levels, not agonist-induced amylase release, in AR42J cells.

机译：Rab3D调节AR42J细胞中的淀粉酶水平，而不是激动剂诱导的淀粉酶释放。

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摘要

Rab3D is a low molecular weight GTP-binding protein that associates with secretory granules in exocrine cells. AR42J cells are derived from rat pancreatic exocrine tumor cells and develop an acinar cell-like phenotype when treated with dexamethasone (Dex). In the present study, we examined the role of Rab3D in Dex-treated AR42J cells. Rab3D expression and localization were analyzed by subcellular fractionation and immunoblotting. The role of Rab3D was examined by overexpressing myc-labeled wild-type-Rab3D and a constitutively active form of Rab3D (Rab3D-Q81L) in AR42J cells. We found that Rab3D is predominantly membrane-associated in AR42J cells and co-localizes with zymogen granules (ZG). Following CCK-8-induced exocytosis, amylase-positive ZGs appeared to move towards the periphery of the cell and co-localization between Rab3D and amylase was less complete when compared to basal conditions. Overexpression of WT, but not mutant Rab3D, resulted in an increase in cellular amylase levels. Overexpression of mutant and WT Rab3D did not affect granule morphology, CCK-8-induced secretion, long-term (48 hr) basal amylase release or granule density. We conclude that Rab3D is not involved in agonist-induced exocytosis in AR42J cells. Instead, Rab3D may regulate amylase content in these cells.
机译：Rab3D是一种低分子量GTP结合蛋白，与外分泌细胞中的分泌颗粒相关。 AR42J细胞衍生自大鼠胰腺外分泌肿瘤细胞，并在用地塞米松（Dex）处理后发展成腺泡细胞样表型。在本研究中，我们检查了Rab3D在Dex处理的AR42J细胞中的作用。通过亚细胞分级分离和免疫印迹分析Rab3D的表达和定位。通过在AR42J细胞中过表达myc标记的野生型Rab3D和Rab3D的组成型活性形式（Rab3D-Q81L）来检查Rab3D的作用。我们发现Rab3D主要是在AR42J细胞中与膜相关，并与酶原颗粒（ZG）共定位。在CCK-8诱导的胞吐作用之后，淀粉酶阳性ZG似乎向细胞外围移动，与基础条件相比，Rab3D和淀粉酶之间的共定位不完全。 WT，但不是突变Rab3D的过表达导致细胞淀粉酶水平的增加。突变体和野生型Rab3D的过表达不影响颗粒形态，CCK-8诱导的分泌，长期（48小时）基础淀粉酶释放或颗粒密度。我们得出结论，Rab3D不参与AR42J细胞中激动剂诱导的胞吐作用。相反，Rab3D可以调节这些细胞中的淀粉酶含量。

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