Reinhardt Annekathrin, Liu Hongyu, Ma Yunxia, Zhou Yongan, Zang Chuanbing, Habbel Jan-Piet, Possinger Kurt, Eucker Jan
Department of Neuropathology, Institute for Pathology, University of Heidelberg, Heidelberg, Germany.
Department of Hematology and Oncology, Benjamin Franklin Campus, Charité University of Medicine Berlin, Berlin, Germany.
Anticancer Res. 2018 May;38(5):2669-2682. doi: 10.21873/anticanres.12509.
BACKGROUND/AIM: One of the major problems in breast cancer treatment is pharmacoresistance. Therefore, exploration of treatment alternatives is of clinical relevance. The present work focused on tumor cell-inhibiting effects of a combination of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and all trans retinoic acid (ATRA) in breast cancer cells.
Breast cancer cell lines (BT-20, BT-474, MDA-MB-231, MDA-MB-436, MDA-MB-453, MCF-7, SKBR3, T47D, ZR-75-1) and the mammary epithelial cell line MCF-10A were treated with TRAIL and ATRA alone and in combination. Cell viability was assessed via 3-(4,5)-dimethylthiahiazo(-z-yl)-3,5-di-phenytetrazoliumromide (MTT) assay, the potential of cell colony formation via clonogenic assay, cell death induction via cell-cycle analysis by fluorescence-activated cell sorting (FACS), terminal deoxynucleotidyltransferase-mediated UTP nick end labeling (TUNEL) assay and Cell death detection ELISA, expression of apoptosis and TRAIL pathway proteins via western blot and cell surface expression of TRAIL receptor 1 (DR4) via FACS analysis.
TRAIL and ATRA evoked synergistic inhibition of breast cancer cell viability based on cytostatic and cytotoxic mechanisms. This correlated with augmented fragmentation of nuclear DNA, up-regulation of TRAIL receptor, down-regulation of cyclin D1 and enhancement of caspase activity. MCF-10A cells were merely slightly susceptible to TRAIL and ATRA.
The cytostatic and cytotoxic effects of the combination of TRAIL and ATRA are tumor cell-selective.
背景/目的：乳腺癌治疗中的主要问题之一是耐药性。因此，探索替代治疗方法具有临床意义。目前的研究聚焦于肿瘤坏死因子相关凋亡诱导配体（TRAIL）与全反式维甲酸（ATRA）联合对乳腺癌细胞的肿瘤细胞抑制作用。
用TRAIL和ATRA单独及联合处理乳腺癌细胞系（BT - 20、BT - 474、MDA - MB - 231、MDA - MB - 436、MDA - MB - 453、MCF - 7、SKBR3、T47D、ZR - 75 - 1）和乳腺上皮细胞系MCF - 10A。通过3 -（4,5）-二甲基噻唑 - 2,5 - 二苯基四氮唑溴盐（MTT）法评估细胞活力，通过克隆形成试验评估细胞集落形成潜力，通过荧光激活细胞分选（FACS）进行细胞周期分析、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）试验和细胞死亡检测ELISA来诱导细胞死亡，通过蛋白质印迹法检测凋亡和TRAIL途径蛋白的表达，并通过FACS分析检测TRAIL受体1（DR4）的细胞表面表达。
基于细胞生长抑制和细胞毒性机制，TRAIL和ATRA对乳腺癌细胞活力产生协同抑制作用。这与核DNA片段增加、TRAIL受体上调、细胞周期蛋白D1下调和半胱天冬酶活性增强相关。MCF - 10A细胞对TRAIL和ATRA仅略有敏感性。
TRAIL与ATRA联合的细胞生长抑制和细胞毒性作用具有肿瘤细胞选择性。
