Ta Na, Huang Xiaoyi, Zheng Kailian, Zhang Yunshuo, Gao Yisha, Deng Lulu, Zhang Bingbing, Jiang Hui, Zheng Jianming
Department of Pathology, Changhai Hospital, Second Military Medical University, Shanghai, China.
Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China.
Cell Physiol Biochem. 2018;51(2):711-728. doi: 10.1159/000495328. Epub 2018 Nov 21.
BACKGROUND/AIMS: MicroRNAs (miRNAs) are a group of non-coding RNAs that play diverse roles in pancreatic carcinogenesis. In pancreatic ductal adenocarcinoma (PDAC), NF-kB is constitutively activated in most patients and is linked to a mutation in KRAS via IkB kinase complex 1 (IKK1, also known as IKKa). We investigated the link between PDAC aggressiveness and miR-1290.
We used miRCURYTM LNA Array and in situ hybridization to investigate candidate miRNAs and validated the findings with PCR. The malignant behavior of cell lines was assessed with Cell Counting Kit-8, colony formation, and Transwell assays. A dual-luciferase reporter assay was used to evaluate the interaction between miR-1290 and IKK1. Protein expression was observed by western blotting.
In this study, 36 miRNAs were dysregulated in high-grade pancreatic intraepithelial neoplasia (PanIN) and PDAC tissues compared with low-grade PanIN tissues. The area under the curve values of miR-1290 and miR-31-5p were 0.829 and 0.848, respectively (95% confidence interval, 0.722-0.936 and 0.749-0.948, both P < 0.001). There was a significant correlation between miR-1290 and histological differentiation (P = 0.029), pT stage (P = 0.006), and lymph node metastasis (P = 0.001). In addition, the in vitro work showed that miR-1290 promoted PDAC cell proliferation, invasion, and migration. Western blotting and the dual-luciferase reporter assay showed that miR-1290 promoted cancer aggressiveness by directly targeting IKK1. The synergist effect of miR-1290 on the proliferation and metastasis of PDAC cells was attenuated and enhanced by IKK1 overexpression and knockdown, respectively. Consistent with the in vitro results, a subcutaneous tumor mouse model showed that miR-1290 functioned as a potent promoter of PDAC in vivo.
MiR-1290 may act as an oncogene by directly targeting the 3'-untranslated region of IKK1, and the miR-1290/IKK1 pathway may prove to be a novel diagnostic and therapeutic target for PDAC.
背景/目的：微小RNA（miRNA）是一类非编码RNA，在胰腺癌发生过程中发挥多种作用。在胰腺导管腺癌（PDAC）中，大多数患者的核因子-κB（NF-κB）持续激活，并通过IkB激酶复合物1（IKK1，也称为IKKa）与KRAS突变相关。我们研究了PDAC侵袭性与miR-1290之间的联系。
我们使用miRCURYTM LNA芯片和原位杂交来研究候选miRNA，并通过聚合酶链反应（PCR）验证结果。使用细胞计数试剂盒-8、集落形成和Transwell实验评估细胞系的恶性行为。采用双荧光素酶报告基因实验评估miR-1290与IKK1之间的相互作用。通过蛋白质印迹法观察蛋白质表达。
在本研究中，与低级别胰腺上皮内瘤变（PanIN）组织相比，高级别PanIN和PDAC组织中有36种miRNA表达失调。miR-1290和miR-31-5p的曲线下面积值分别为0.829和0.848（95%置信区间，0.722 - 0.936和0.749 - 0.948，均P < 0.001）。miR-1290与组织学分化（P = 0.029）、pT分期（P = 0.006）和淋巴结转移（P = 0.001）之间存在显著相关性。此外，体外实验表明miR-1290促进PDAC细胞增殖、侵袭和迁移。蛋白质印迹法和双荧光素酶报告基因实验表明，miR-1290通过直接靶向IKK1促进癌症侵袭性。IKK1过表达和敲低分别减弱和增强了miR-1290对PDAC细胞增殖和转移的协同作用。与体外结果一致，皮下肿瘤小鼠模型表明miR-1290在体内是PDAC的有效促进因子。
MiR-1290可能通过直接靶向IKK1的3'-非翻译区发挥癌基因作用，miR-1290/IKK1通路可能成为PDAC新的诊断和治疗靶点。
