Ohana Joel, Sandler Uziel, Kass Gideon, Stemmer Salomon M, Devary Yoram
Immune System Key (ISK) Ltd., Jerusalem 9746009, Israel.
Department of Bio-Informatics, Lev Academic Center (JCT), Jerusalem 91160, Israel.
Mol Clin Oncol. 2017 Dec;7(6):991-999. doi: 10.3892/mco.2017.1453. Epub 2017 Oct 17.
dTCApFs (Nerofe™) is a 14-amino acid derivative of a longer hormone peptide, tumor-cells apoptosis factor (TCApF), which enters the cells through the T1/ST2 receptor. In the present study, the mechanism of action (MOA) of dTCApFs as an anticancer agent was investigated. Experiments were performed in pancreatic cancer cell lines, and immunofluorescent staining demonstrated that dTCApFs is located in the Golgi apparatus of treated cells. It was also demonstrated in pancreatic, breast and ovarian cell lines that dTCApFs treatment led to Golgi structural changes, loss of Golgi function, and molecular effects associated with endoplasmic reticulum (ER) stress, such as increased levels of C/EBP homologous protein, binding immunoglobulin protein (BiP), phosphorylated inositol-requiring enzyme 1 (pIRE1), and increased phosphorylation of eukaryotic translation initiation factor 2α, and to the generation of reactive oxygen species, which was attenuated by ER stress inhibitors. Moreover, in these cell lines, long-term exposure to dTCApFs led to downregulation of spliced X-box-binding protein 1, which is an ER stress repair mechanism gene, downregulation of the Golgi anti-apoptotic protein, and reduced cell viability. studies using murine xenograft models of human pancreatic cancer verified the cell culture findings by demonstrating structural changes in the ER/Golgi and increased levels of pIRE1and BiP in dTCApFs-treated mice vs. the controls. Finally, human tissue samples from a patient who received dTCApFs for 11 months in a clinical trial were analyzed, and an increase was observed in the number of cells expressing pIRE1 and BiP post-treatment. In conclusion, we herein report a novel MOA for an anticancer agent involving triggering of apoptosis through induction of opposite effects: ER stress and downregulation of the ER stress repair mechanism. These findings provide the framework for the clinical evaluation of dTCApFs.
十二碳十四酸肿瘤细胞凋亡因子（Nerofe™）是一种由更长的激素肽——肿瘤细胞凋亡因子（TCApF）衍生而来的14个氨基酸的肽段，它通过T1/ST2受体进入细胞。在本研究中，对十二碳十四酸肿瘤细胞凋亡因子作为抗癌剂的作用机制进行了研究。实验在胰腺癌细胞系中进行，免疫荧光染色显示十二碳十四酸肿瘤细胞凋亡因子位于经处理细胞的高尔基体中。在胰腺、乳腺和卵巢癌细胞系中还证实，十二碳十四酸肿瘤细胞凋亡因子处理导致高尔基体结构改变、高尔基体功能丧失以及与内质网（ER）应激相关的分子效应，如C/EBP同源蛋白、结合免疫球蛋白蛋白（BiP）、磷酸化肌醇需求酶1（pIRE1）水平升高，真核翻译起始因子2α磷酸化增加，以及活性氧的产生，而内质网应激抑制剂可减弱这些效应。此外，在这些细胞系中，长期暴露于十二碳十四酸肿瘤细胞凋亡因子会导致剪接X盒结合蛋白1（一种内质网应激修复机制基因）下调、高尔基体抗凋亡蛋白下调以及细胞活力降低。使用人胰腺癌小鼠异种移植模型的研究通过证明与对照组相比，经十二碳十四酸肿瘤细胞凋亡因子处理的小鼠内质网/高尔基体结构改变以及pIRE1和BiP水平升高，验证了细胞培养结果。最后，对一名在临床试验中接受十二碳十四酸肿瘤细胞凋亡因子治疗11个月的患者的人体组织样本进行分析，发现治疗后表达pIRE1和BiP的细胞数量增加。总之，我们在此报告了一种抗癌剂的新作用机制，即通过诱导相反的效应引发凋亡：内质网应激和内质网应激修复机制下调。这些发现为十二碳十四酸肿瘤细胞凋亡因子的临床评估提供了框架。
