Matuszcak Christiane, Lindner Kirsten, Eichelmann Ann-Kathrin, Hussey Damian J, Haier Jörg, Hummel Richard
University Cancer Centre Hamburg (UCCH), University Hospital of Hamburg-Eppendorf, Martinistr. 52 (O24), 20246 Hamburg, Germany.
Department of Surgery, University of Schleswig-Holstein, Lübeck, Germany.
Surg Oncol. 2018 Sep;27(3):392-401. doi: 10.1016/j.suronc.2018.04.001. Epub 2018 Apr 21.
Incidence of esophageal adenocarcinoma (EAC) increased significantly over the last decades. Lack of response to chemotherapy is a major problem in the treatment of this disease. This study aims to assess the biological relevance of characteristic microRNA profiles of chemotherapy resistant EAC cells with regards to response to chemotherapy and biological behavior.
We selected 3 microRNAs from characteristic microRNA profiles of resistant EAC (miR-27b-3p, miR-200b-3p, and miR-148a-3p). Expression of microRNAs was modified in 6 EAC cell lines. Effects on chemotherapy, adhesion, migration, apoptosis and cell cycle were assessed using standard assays. Target analyses were performed using Western Blot and Luciferase techniques.
MiR-27b-3p significantly sensitized cells to 5FU and Cisplatin in 83% respectively in 33% of cell lines, miR-148a-3p in 67% respectively 33% of cases. MiR-200b-3p increased sensitivity only towards 5FU in 50% of cases. Co-transfections with miR-27b-3p/miR-148a-3p showed an additive effect on response to chemotherapy in 50% of cases. Upregulation of miR-148a-3p reduced protein expression levels of DNMT-1, MSK-1, Bcl-2 and Bim, and miR-27b upregulation led to downregulation of Sp1 and PPARy proteins implicating a potential negative post-transcriptional control via the respective microRNAs. Finally, we were able to confirm Bcl-2 for the first time as direct target of miR-148a-3p in EAC.
This study demonstrates that specific microRNA profiles of chemotherapy resistant EAC in fact determine their response to chemotherapy and biological behavior. Our data further show that microRNA-mediated regulation of chemotherapy resistance is complex, and several microRNAs seem to "co-operate" at various steps within a broad number of pathways what fits very well to our recently proposed understanding of microRNA-mediated regulation as function of cellular functional complexes. These data highlight the promising potential of microRNAs to predict or monitor treatment response to chemotherapy in EAC, and to potentially modulate tumor biology in a therapeutic approach.
在过去几十年中，食管腺癌（EAC）的发病率显著上升。对化疗无反应是该疾病治疗中的一个主要问题。本研究旨在评估化疗耐药EAC细胞特征性微小RNA谱在化疗反应和生物学行为方面的生物学相关性。
我们从耐药EAC的特征性微小RNA谱中选择了3种微小RNA（miR-27b-3p、miR-200b-3p和miR-148a-3p）。在6种EAC细胞系中改变微小RNA的表达。使用标准检测方法评估对化疗、黏附、迁移、凋亡和细胞周期的影响。使用蛋白质印迹法和荧光素酶技术进行靶点分析。
MiR-27b-3p分别在33%和83%的细胞系中使细胞对5-氟尿嘧啶和顺铂显著敏感，miR-148a-3p分别在33%和67%的病例中显示出同样效果。MiR-200b-3p仅在50%的病例中增加了对5-氟尿嘧啶的敏感性。miR-27b-3p/miR-14-8a-3p共转染在50%的病例中对化疗反应显示出相加效应。miR-148a-3p的上调降低了DNMT-1、MSK-1、Bcl-2和Bim的蛋白表达水平，miR-27b的上调导致Sp1和PPARγ蛋白的下调，这暗示了通过各自的微小RNA进行潜在的负转录后调控。最后，我们首次能够证实Bcl-2是EAC中miR-148a-3p的直接靶点。
本研究表明，化疗耐药EAC的特定微小RNA谱实际上决定了它们对化疗的反应和生物学行为。我们的数据进一步表明，微小RNA介导的化疗耐药调控是复杂的，几种微小RNA似乎在众多途径的不同步骤中“协同作用”，这与我们最近提出的将微小RNA介导的调控理解为细胞功能复合物功能的观点非常契合。这些数据突出了微小RNA在预测或监测EAC化疗反应以及在治疗方法中潜在调节肿瘤生物学方面的巨大潜力。
