Lantto Tiina A, Laakso Into, Dorman H J Damien, Mauriala Timo, Hiltunen Raimo, Kõks Sulev, Raasmaja Atso
Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Post Office Box 56 (Viikinkaari 5E), Helsinki 00014, Finland.
Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, 19 Ravila Street, Tartu 50411, Estonia.
Int J Mol Sci. 2016 Jul 13;17(7):1113. doi: 10.3390/ijms17071113.
Plant phenolics have shown to activate apoptotic cell death in different tumourigenic cell lines. In this study, we evaluated the effects of juniper berry extract (Juniperus communis L.) on p53 protein, gene expression and DNA fragmentation in human neuroblastoma SH-SY5Y cells. In addition, we analyzed the phenolic composition of the extract. We found that juniper berry extract activated cellular relocalization of p53 and DNA fragmentation-dependent cell death. Differentially expressed genes between treated and non-treated cells were evaluated with the cDNA-RDA (representational difference analysis) method at the early time point of apoptotic process when p53 started to be activated and no caspase activity was detected. Twenty one overexpressed genes related to cellular stress, protein synthesis, cell survival and death were detected. Interestingly, they included endoplasmic reticulum (ER) stress inducer and sensor HSPA5 and other ER stress-related genes CALM2 and YKT6 indicating that ER stress response was involved in juniper berry extract mediated cell death. In composition analysis, we identified and quantified low concentrations of fifteen phenolic compounds. The main groups of them were flavones, flavonols, phenolic acids, flavanol and biflavonoid including glycosides of quercetin, apigenin, isoscutellarein and hypolaetin. It is suggested that juniper berry extract induced the p53-associated apoptosis through the potentiation and synergism by several phenolic compounds.
植物酚类物质已被证明能在不同的致瘤细胞系中激活凋亡性细胞死亡。在本研究中，我们评估了杜松子提取物（杜松）对人神经母细胞瘤SH-SY5Y细胞中p53蛋白、基因表达和DNA片段化的影响。此外，我们分析了提取物的酚类成分。我们发现杜松子提取物激活了p53的细胞重新定位和依赖DNA片段化的细胞死亡。在凋亡过程的早期时间点，当p53开始被激活且未检测到半胱天冬酶活性时，用cDNA-RDA（代表性差异分析）方法评估处理细胞和未处理细胞之间差异表达的基因。检测到21个与细胞应激、蛋白质合成、细胞存活和死亡相关的过表达基因。有趣的是，它们包括内质网（ER）应激诱导剂和传感器HSPA5以及其他ER应激相关基因CALM2和YKT6，这表明ER应激反应参与了杜松子提取物介导的细胞死亡。在成分分析中，我们鉴定并定量了低浓度的15种酚类化合物。它们的主要类别是黄酮、黄酮醇、酚酸、黄烷醇和双黄酮，包括槲皮素、芹菜素、异灯盏乙素和次黄嘌呤素的糖苷。提示杜松子提取物通过几种酚类化合物的增强和协同作用诱导p53相关的细胞凋亡。
