Flammang Isabelle, Reese Moritz, Ströse Anda J., Yang Zixuan, Eble Johannes A, Dhayat Sameer A
Department of General, Visceral and Transplantation Surgery, University Hospital Muenster, Albert-Schweitzer-Campus 1 (W1), 48149 Muenster, Germany.
Department of Physiological Chemistry and Pathobiochemistry, University of Muenster, Waldeyerstrasse 15, 48149 Muenster, Germany.
Cancers (Basel). 2020 Jun 25;12(6):1693. doi: 10.3390/cancers12061693.
Pancreatic ductal adenocarcinoma (PDAC) is characterized by fast tumor progression and diagnosis at advanced, inoperable stages. Previous studies could demonstrate an involvement of miR-192-5p in epigenetic regulation of visceral carcinomas. Due to contradictory results, however, the clinical utility of miR-192-5p in PDAC has yet to be determined. MiR-192-5p expression was analyzed by RT-qRT-PCR in human PDAC and benign tissue ( = 78), blood serum ( = 81) and serum exosomes ( = 74), as well as in PDAC cell lines ( = 5), chemoresistant cell clones ( = 2), and pancreatic duct cell line H6c7. Analysis of EMT-associated (epithelial-to-mesenchymal transition) proteins was performed by immunohistochemistry and Western blot. MiR-192-5p was deregulated in PDAC as compared to healthy controls (HCs), with downregulation in macrodissected tissue ( < 0.001) and upregulation in blood serum of PDAC UICC (Union for International Cancer Control) stage IV ( = 0.016) and serum exosomes of PDAC UICC stages II to IV ( < 0.001). MiR-192-5p expression in tumor tissue was significantly lower as compared to corresponding peritumoral tissue (PDAC UICC stage II: < 0.001; PDAC UICC stage III: = 0.024), while EMT markers ZEB1 and ZEB2 were more frequently expressed in tumor tissue as compared to peritumoral tissue, HCs, and chronic pancreatitis. Tissue-derived (AUC of 0.86; < 0.0001) and exosomal (AUC of 0.83; = 0.0004) miR-192-5p could differentiate between PDAC and HCs with good accuracy. Furthermore, high expression of miR-192-5p in PDAC tissue of curatively resected PDAC patients correlated with prolonged overall and recurrence-free survival in multivariate analysis. In vitro, miR-192-5p was downregulated in gemcitabine-resistant cell clones of AsPC-1 ( = 0.029). Transient transfection of MIA PaCa-2 cells with miR-192-5p mimic resulted in downregulation of ZEB2. MiR-192-5p seems to possess a tumor-suppressive role and high potential as a diagnostic and prognostic marker in PDAC.
胰腺导管腺癌（PDAC）的特征是肿瘤进展迅速，且多在晚期、无法手术切除的阶段被诊断出来。先前的研究表明miR-192-5p参与了内脏癌的表观遗传调控。然而，由于结果相互矛盾，miR-192-5p在PDAC中的临床应用尚未确定。通过逆转录定量聚合酶链反应（RT-qRT-PCR）分析了miR-192-5p在人PDAC组织和良性组织（n = 78）、血清（n = 81）和血清外泌体（n = 74）中的表达，以及在PDAC细胞系（n = 5）、化疗耐药细胞克隆（n = 2）和胰腺导管细胞系H6c7中的表达。通过免疫组织化学和蛋白质印迹法分析上皮-间质转化（EMT）相关蛋白。与健康对照（HCs）相比，PDAC中miR-192-5p失调，在大块解剖组织中下调（P < 0.001），在PDAC国际癌症控制联盟（UICC）IV期血清中上调（P = 0.016），在PDAC UICC II至IV期血清外泌体中上调（P < 0.001）。肿瘤组织中miR-192-5p的表达明显低于相应的瘤旁组织（PDAC UICC II期：P < 0.001；PDAC UICC III期：P = 0.024），而EMT标志物锌指蛋白E盒结合因子1（ZEB1）和锌指蛋白E盒结合因子2（ZEB2）在肿瘤组织中的表达比瘤旁组织、HCs和慢性胰腺炎中更频繁。组织来源的miR-192-5p（曲线下面积[AUC]为0.86；P < 0.0001）和外泌体来源的miR-192-5p（AUC为0.83；P = 0.0004）能够很好地区分PDAC和HCs。此外，在多变量分析中，根治性切除的PDAC患者的PDAC组织中miR-192-5p的高表达与总体生存期和无复发生存期的延长相关。在体外，阿帕奇-1（AsPC-1）吉西他滨耐药细胞克隆中miR-192-5p下调（P = 0.029）。用miR-192-5p模拟物瞬时转染MIA PaCa-2细胞导致ZEB2下调。miR-192-5p似乎具有肿瘤抑制作用，在PDAC中作为诊断和预后标志物具有很高的潜力。
