人胰腺导管腺癌细胞中功能性瞬时受体电位香草酸亚家族成员8通道的特性分析

Characterization of functional transient receptor potential melastatin 8 channels in human pancreatic ductal adenocarcinoma cells.

作者信息

Cucu Dana, Chiritoiu Gabriela, Petrescu Stefana, Babes Alexandru, Stanica Luciana, Duda Dan G, Horii Akira, Dima Simona Olimpia, Popescu Irinel

机构信息

From the *Center of Digestive Disease and Liver Transplantation, Fundeni Clinical Institute; †Department of Molecular and Cell Biology, Romanian Academy Institute of Biochemistry; ‡Department of Anatomy, Physiology, and Biophysics, Faculty of Biology, University of Bucharest, Bucharest, Romania; §Department of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA; and ∥Division of Molecular Pathology, Department of Pathology, Tohoku University Graduate School of Medicine, Sendai, Japan.

出版信息

Pancreas. 2014 Jul;43(5):795-800. doi: 10.1097/MPA.0000000000000106.

DOI:10.1097/MPA.0000000000000106

PMID:24658318

Abstract

OBJECTIVE

Recently, the transient receptor potential melastatin 8 (TRPM8) channel has emerged as a putative biomarker for pancreatic ductal adenocarcinoma (PDA). This study aimed to evaluate the expression of TRPM8 and its modulation by specific agonists and antagonists in PDA cells.

METHODS

We examined the protein expression of TRPM8 in 3 different PDA cell lines and compared it with a nontumoral epithelial cell line of human pancreatic origin using Western blotting and immunocytochemical analysis. To assess the function of TRPM8 channels, we measured the TRPM8 currents in whole-cell mode of the patch clamp technique. To explore the putative involvement of TRPM8 in cell migration, we investigated the motility of PDA cells using the scratch-wound assay.

RESULTS

Pancreatic ductal adenocarcinoma cells express functional plasma membrane TRPM8 channels, which are responsive after exposure to agonists (menthol and icilin) and antagonists N-(3-aminopropyl)-2-{[(3-methylphenyl) methyl]oxy}-N-(2-thienylmethyl)benzamide hydrochloride salt. The silencing of TRPM8 expression by small interfering RNA augments the migration of PDA cells. Conversely, the activated form of TRPM8 inhibits PDA cell motility.

CONCLUSIONS

An unglycosylated TRPM8 protein is expressed and is functional in the membrane of PDA cells. Transient receptor potential melastatin 8 inhibits the migration of PDA cells, suggesting a putative role as a biomarker or target for this channel for PDA therapy.

摘要

目的

最近，瞬时受体电位香草酸亚型8（TRPM8）通道已成为胰腺导管腺癌（PDA）的一种潜在生物标志物。本研究旨在评估TRPM8在PDA细胞中的表达及其受特异性激动剂和拮抗剂的调节情况。

方法

我们使用蛋白质印迹法和免疫细胞化学分析法，检测了3种不同PDA细胞系中TRPM8的蛋白表达，并将其与源自人胰腺的非肿瘤上皮细胞系进行比较。为评估TRPM8通道的功能，我们采用膜片钳技术的全细胞模式测量TRPM8电流。为探究TRPM8在细胞迁移中的潜在作用，我们使用划痕试验研究了PDA细胞的运动性。

结果

胰腺导管腺癌细胞表达功能性质膜TRPM8通道，在暴露于激动剂（薄荷醇和异冰片）和拮抗剂盐酸N-（3-氨丙基）-2-{[（3-甲基苯基）甲基]氧基}-N-（2-噻吩基甲基）苯甲酰胺后会产生反应。小干扰RNA使TRPM8表达沉默会增强PDA细胞的迁移。相反，TRPM8的激活形式会抑制PDA细胞的运动性。