Du Wenwen, Liu Ting, Zhang Yang, Zeng Yuanyuan, Zhu Jianjie, Tang Haicheng, Liu Zeyi, Huang Jian-An
Department of Respiratory Medicine, the First Affiliated Hospital of Soochow University, Suzhou, 215006, China.
Suzhou Key Laboratory for Respiratory Diseases, Suzhou, 215006, China.
J Cancer. 2020 Feb 19;11(9):2610-2620. doi: 10.7150/jca.39884. eCollection 2020.
Lung cancer is the most common malignancy with poor 5-year survival among men and women. Previous studies have shown that CPNE1 is up-regulated in non-small cell lung cancer (NSCLC). However, whether and how CPNE1 expression varies between different subtypes of NSCLC remains less understood. Bioinformatical analysis and GSE19188 were selected to confirm CPNE1 expression in different subtypes of NSCLC. Four microRNA prediction websites and GSE53883, GSE43000 were used to evaluate the possible targeting microRNAs. Kaplan-Meier survival curves were drawn based on Tumor Lung Bild -114 dataset using R2, UCSC Xena browser or linkedomics platform. Furthermore, we verified our prediction via qRT-PCR, and western blot and luciferase reporter assays. we demonstrated that higher CPNE1 expression was associated with poorer survival in NSCLC patients. Moreover, among the different subtypes, patients with squamous cell lung cancer (SCC) exhibited higher level of CPNE1 expression, as well as substantially poorer survival. MiR-195-5p was down-regulated in NSCLC tissues. Interestingly, SCC patients showed lower miR-195-5p expression compared to patients with lung adenocarcinoma (ADC). In addition, functional assays proved that miR-195-5p overexpression inhibited the proliferation, migration, and invasion of NSCLC-derived cells by directly targeting CPNE1. Pathway analysis showed decreased expression of p-AKT, p-Erk, and Snail after transfection with miR-195-5p mimics in both lung adenocarcinoma and squamous cell lines. Our findings suggested that miR-195-5p regulation contributed to the differential expression of CPNE1 in NSCLC subtypes.
肺癌是男性和女性中最常见的恶性肿瘤，5年生存率较低。先前的研究表明，CPNE1在非小细胞肺癌（NSCLC）中上调。然而，CPNE1在不同亚型的NSCLC中的表达差异以及其具体机制仍不清楚。本研究通过生物信息学分析和GSE19188数据集来确认CPNE1在不同亚型NSCLC中的表达。使用四个microRNA预测网站以及GSE53883、GSE43000数据集来评估可能靶向CPNE1的microRNA。基于Tumor Lung Bild -114数据集，使用R2、UCSC Xena浏览器或linkedomics平台绘制Kaplan-Meier生存曲线。此外，我们通过qRT-PCR、蛋白质免疫印迹和荧光素酶报告基因检测验证了我们的预测。我们证明，NSCLC患者中CPNE1表达越高，生存率越低。此外，在不同亚型中，肺鳞状细胞癌（SCC）患者的CPNE1表达水平更高，生存率也显著更低。MiR-195-5p在NSCLC组织中表达下调。有趣的是，与肺腺癌（ADC）患者相比，SCC患者的miR-195-5p表达更低。此外，功能实验证明，miR-195-5p过表达通过直接靶向CPNE1抑制NSCLC来源细胞的增殖、迁移和侵袭。通路分析显示，在肺腺癌和鳞状细胞系中转染miR-195-5p模拟物后，p-AKT、p-Erk和Snail的表达降低。我们的研究结果表明，miR-195-5p调控导致了CPNE1在NSCLC亚型中的差异表达。
