Skrzypski Marek, Kołodziejski Paweł A, Mergler Stefan, Khajavi Noushafarin, Nowak Krzysztof W, Strowski Mathias Z
Department of Animal Physiology and Biochemistry, Poznań University of Life Sciences, 60-637 Poznań, Poland.
Department of Animal Physiology and Biochemistry, Poznań University of Life Sciences, 60-637 Poznań, Poland Department of Hepatology and Gastroenterology and Interdisciplinary Centre of Metabolism: Endocrinology, Diabetes and Metabolism, Charité-University Medicine Berlin, 13353 Berlin, Germany.
Biosci Rep. 2016 Aug 24;36(4). doi: 10.1042/BSR20160106. Print 2016 Aug.
Highly Ca(2+) permeable receptor potential channel vanilloid type 6 (TRPV6) modulates a variety of biological functions including calcium-dependent cell growth and apoptosis. So far, the role of TRPV6 in controlling growth of pancreatic neuroendocrine tumour (NET) cells is unknown. In the present study, we characterize the expression of TRPV6 in pancreatic BON-1 and QGP-1 NET cells. Furthermore, we evaluate the impact of TRPV6 on intracellular calcium, the activity of nuclear factor of activated T-cells (NFAT) and proliferation of BON-1 cells. TRPV6 expression was assessed by real-time PCR and Western blot. TRPV6 mRNA expression and protein production were down-regulated by siRNA. Changes in intracellular calcium levels were detected by fluorescence calcium imaging (fura-2/AM). NFAT activity was studied by NFAT reporter assay; cell proliferation by bromodeoxyuridine (BrdU), MTT and propidium iodine staining. TRPV6 mRNA and protein are present in BON-1 and QGP-1 NET-cells. Down-regulation of TRPV6 attenuates BON-1 cell proliferation. TRPV6 down-regulation is associated with decreased Ca(2+) response pattern and reduced NFAT activity. In conclusion, TRPV6 is expressed in pancreatic NETs and modulates cell proliferation via Ca(2+)-dependent mechanism, which is accompanied by NFAT activation.
高钙（Ca²⁺）通透性香草酸受体亚型6（TRPV6）调节多种生物学功能，包括钙依赖性细胞生长和凋亡。到目前为止，TRPV6在控制胰腺神经内分泌肿瘤（NET）细胞生长中的作用尚不清楚。在本研究中，我们对TRPV6在胰腺BON-1和QGP-1 NET细胞中的表达进行了表征。此外，我们评估了TRPV6对细胞内钙、活化T细胞核因子（NFAT）活性以及BON-1细胞增殖的影响。通过实时PCR和蛋白质印迹法评估TRPV6的表达。通过小干扰RNA（siRNA）下调TRPV6 mRNA表达和蛋白质产生。通过荧光钙成像（fura-2/AM）检测细胞内钙水平的变化。通过NFAT报告基因检测研究NFAT活性；通过溴脱氧尿苷（BrdU）、MTT和碘化丙啶染色检测细胞增殖。TRPV6 mRNA和蛋白质存在于BON-1和QGP-1 NET细胞中。TRPV6的下调减弱了BON-1细胞的增殖。TRPV6下调与Ca²⁺反应模式降低和NFAT活性降低相关。总之，TRPV6在胰腺NET中表达，并通过Ca²⁺依赖性机制调节细胞增殖，这伴随着NFAT的激活。
