Kanno Shinichi, Nosho Katsuhiko, Ishigami Keisuke, Yamamoto Itaru, Koide Hideyuki, Kurihara Hiroyoshi, Mitsuhashi Kei, Shitani Masahiro, Motoya Masayo, Sasaki Shigeru, Tanuma Tokuma, Maguchi Hiroyuki, Hasegawa Tadashi, Kimura Yasutoshi, Takemasa Ichiro, Shinomura Yasuhisa, Nakase Hiroshi
Department of Gastroenterology and Hepatology, Sapporo Medical University School of Medicine, Sapporo 006-0811, Japan.
Department of Gastroenterology, Teine Keijinkai Hospital, Sapporo, Japan.
Carcinogenesis. 2017 Apr 1;38(4):425-431. doi: 10.1093/carcin/bgx013.
Pancreatic cancer is a highly aggressive malignancy, with <50% patients surviving beyond 6 months after the diagnosis, and thus, there is an urgent need to explore new diagnostic and therapeutic approaches for this disease. Therefore, we conducted microRNA (miRNA) array analysis to detect miRNA molecules potentially associated with pancreatic cancer malignancy. To assess the identified miRNAs, we performed quantitative reverse transcription-PCR on 248 pancreatic ductal adenocarcinomas (UICC stage II). We also examined miRNA expression [microRNA-21 (miR-21) and microRNA-31 (miR-31)] and epigenetic alterations, including CpG island methylator phenotype (CIMP), potentially associated with the identified miRNAs. For functional analysis, we conducted proliferation and invasion assays using a pancreatic cancer cell line. miRNA array analysis revealed that microRNA-196b (miR-196b) was the most up-regulated miRNA in pancreatic cancer tissues compared with normal pancreatic duct cells. High miR-196b expression was associated with miR-21 (P = 0.0025) and miR-31 (P = 0.0001) expression. It was also related to poor prognosis in the multivariate analysis using overall survival (hazard ratio: 1.66; 95% confidence interval: 1.09-2.54; P = 0.019). Functional analysis demonstrated that miR-196b inhibitor decreased cell proliferation and that miR-196b mimic promoted cancer cell invasion. In conclusion, a significant association of high miR-196b expression with poor prognosis was observed in pancreatic cancer. Our data also revealed that miR-196b played an oncogenic role and that the transfection of the miR-196b inhibitor had an anti-tumour effect in the pancreatic cancer cell line. These results suggest that miR-196b is a promising diagnostic biomarker and therapeutic target in pancreatic cancer.
胰腺癌是一种侵袭性很强的恶性肿瘤，诊断后存活超过6个月的患者不到50%，因此，迫切需要探索针对这种疾病的新诊断和治疗方法。因此，我们进行了微小RNA（miRNA）芯片分析，以检测可能与胰腺癌恶性程度相关的miRNA分子。为了评估鉴定出的miRNA，我们对248例胰腺导管腺癌（UICC II期）进行了定量逆转录PCR。我们还检测了miRNA表达[微小RNA-21（miR-21）和微小RNA-31（miR-31）]以及表观遗传改变，包括可能与鉴定出的miRNA相关的CpG岛甲基化表型（CIMP）。为了进行功能分析，我们使用胰腺癌细胞系进行了增殖和侵袭试验。miRNA芯片分析显示，与正常胰腺导管细胞相比，微小RNA-196b（miR-196b）是胰腺癌组织中上调最明显的miRNA。高miR-196b表达与miR-21（P = 0.0025）和miR-31（P = 0.0001）表达相关。在使用总生存期的多变量分析中，它也与预后不良有关（风险比：1.66；95%置信区间：1.09 - 2.54；P = 0.019）。功能分析表明，miR-196b抑制剂可降低细胞增殖，而miR-196b模拟物可促进癌细胞侵袭。总之，在胰腺癌中观察到高miR-196b表达与预后不良显著相关。我们的数据还显示，miR-196b发挥致癌作用，并且转染miR-196b抑制剂在胰腺癌细胞系中具有抗肿瘤作用。这些结果表明，miR-196b是胰腺癌中一种有前景的诊断生物标志物和治疗靶点。
