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Bing Han,1,2,* Jiawei Huang,3,* Zhenjie Yang,1 Jiaqi Zhang,3 Xiaomin Wang,3 Ning Xu,3 Haining Meng,3 Junyu Wu,3 Qiao Huang,3 Xi Yang,3 Ruowu Shen,3 Chuandong Sun1 1Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qingdao University, Qingdao City 266003, Shandong Province, People’s Republic of China; 2Asian Liver Center, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA; 3Department of Special Medicine, School of Basic Medical College, Qingdao University, Qingdao City 266021, Shandong Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Chuandong SunDepartment of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qingdao University, No. 16 Jiangsu Road, Qingdao City 266003, Shandong Province, People’s Republic of ChinaTel +86 18661802672Email sunchuandong@hotmail.comRuowu ShenDepartment of Special Medicine, School of Basic Medical College, Qingdao University, Qingdao City 266021, Shandong Province, People’s Republic of ChinaTel +86 18661785993Email shenruowu@aliyun.comPurpose: To explore the effect of miR-449a inhibits migration and invasion by targeting Notch1 and regulating epithelial–mesenchymal transition (EMT) in hepatocellular carcinoma (HCC), and further study on the molecular mechanism.Patients and methods: The expression of miR-449a and Notch1 in HCC cells and tissues was detected by qRT-PCR. The HCC cell line HCCLM3 and SMMC-7721 were transfected with miR-449a. The invasion and migration of HCC cell lines were detected by transwell assay and wound healing assay. The Notch pathway and EMT related protein were detected with Western Blotting. The specific binding site of mir-449a on notch1 gene was detected by luciferase assay.Results: We found the expression of miR-449a was related to short-term recurrence of hepatocellular carcinoma after hepatectomy and acted as independent risk factors of DFS and OS. The expression of miR-449a decreased in tumor tissues and HCC cell lines, but the expression of Notch1 increased. The overexpressed miR-449a promoted the invasiveness in vitro by regulating EMT via Notch pathway. Mechanically, miR-449a inhibited the translation of Notch1 protein by binding to 3ʹ UTR of its mRNA directly.Conclusion: miR-449a is short-term recurrence-related miRNA and inhibits the invasion and metastasis ability of HCC cells by regulating EMT via Notch pathway. miR-449a may be a new effective therapeutic target for HCC.Keywords: signal pathway, microRNA, epithelial-mesenchymal transition, bioinformatics

Haining Meng,1 Qiao Huang,1 Xijin Zhang,1 Jiawei Huang,1 Ruowu Shen,1 Bei Zhang21Department of Special Medicine, School of Basic Medical College, Qingdao University, Qingdao 266021, People’s Republic of China; 2Department of Immunology, School of Basic Medical College, Qingdao University, Qingdao 266021, People’s Republic of ChinaBackground: MicroRNAs (miRNAs) are non-coding small RNAs that have been shown to play a key role in the development of many tumors. However, its specific mechanism of action in non-small cell lung cancer (NSCLC) is not very clear.Purpose: This study was to identify the effect of miRNA-449a on NSCLC invasion and migration.Methods: We used quantitative real-time PCR experiments to demonstrate that miRNA-449a is down-regulated in NSCLC tissues and cell lines. We also used the Transwell assay to detect cell invasion and migration, and the Western Blot assay was used to detect protein expression. The dual luciferase assay was used to detect the targeting relationship between miR-449a and ADisintegrin And Metalloproteinases 10 (ADAM10).Results: Our experiments demonstrated that miRNA-449a was down-regulated in NSCLC tissues and cell lines. When miRNA-449a was up-regulated in NSCLC cells, the invasion and migration ability of the cells was weakened, and the expression of ADAM10 was decreased. After down-regulation of miRNA-449a, the cell’s invasion and migration ability was enhanced, and the expression of ADAM10 was increased. Through dual luciferase assays, we also found that miRNA-449a can target ADAM10 to delay the progression of epithelial-mesenchymal transition (EMT) and inhibit invasion and migration.Conclusion: Our experiments demonstrated that miRNA-449a acted as a tumor suppressor gene through inhibiting the expression of ADAM10 in NSCLC.Keywords: miR-449a, non-small cell lung cancer, NSCLC, ADAM10, epithelial-mesenchymal transition, EMT