miR-146a-5p mediates epithelial-mesenchymal transition of oesophageal squamous cell carcinoma via targeting Notch2.
BACKGROUND
Our previous study found that dysregulated microRNA-146a-5p (miR-146a-5p) is involved in oesophageal squamous cell cancer (ESCC) proliferation. This article aimed to evaluate its detailed mechanisms in ESCC epithelial-mesenchymal transition (EMT) progression.


METHODS
Invasion assay, qRT-PCR and western blotting were used to validate the roles of miR-146a-5p and Notch2 in EMT progression. miRNA target gene prediction databases and dual-luciferase reporter assay were used to validate the target gene.


RESULTS
miR-146a-5p inhibitor led to increase of invaded ESCC cells, while miR-146a-5p mimics inhibited invasion ability of ESCC cells. Protein level of E-cadherin decreased, whereas those of Snail and Vimentin increased in the anti-miR-146a-5p group, which demonstrated that miR-146a-5p inhibits EMT progression of ESCC cells. miRNA target gene prediction databases indicated the potential of Notch2 as a direct target gene of miR-146a-5p and dual-luciferase reporter assay validated it. Importantly, shRNA-Notch2 restrained EMT and partially abrogated the inhibiting effects of miR-146a-5p on EMT progression of ESCC cells.


CONCLUSIONS
miR-146a-5p functions as a tumour-suppressive miRNA targeting Notch2 and inhibits the EMT progression of ESCC.
Department of Radiation Oncology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Cong
C
Wang
Department of Gastroenterology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Wenxue
W
Zhang
Department of Hematology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Lin
L
Zhang
Department of Radiation Oncology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Xuan
X
Chen
Department of Imaging, Shandong Medical College, Jinan, Shandong, China.
Fang
F
Liu
Department of Medical Genetics, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences-School of Basic Medicine Peking Union Medical College, Beijing, China.
Jing
J
Zhang
Department of Radiation Oncology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Shanghui
S
Guan
Department of Radiation Oncology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Yi
Y
Sun
Department of Radiation Oncology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Pengxiang
P
Chen
Department of Laboratory Medicine, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Ding
D
Wang
Department of Radiation Oncology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Effat
E
Un Nesa
Department of Radiation Oncology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
Yufeng
Y
Cheng
Department of Laboratory Medicine, and the Keenan Research Centre for Biomedical Science at the Li Ka Shing Knowledge Institute, St. Michael's Hospital, Toronto, Canada.
George M
GM
Yousef
D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology*; D004938:Esophageal Neoplasms / Q000473:pathology*; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D051883:Receptor, Notch2 / Q000502:physiology*
C542542:MIRN146 microRNA, human; D035683:MicroRNAs; C497764:NOTCH2 protein, human; D051883:Receptor, Notch2