MicroRNA-145 Inhibits Cell Migration and Invasion and Regulates Epithelial-Mesenchymal Transition (EMT) by Targeting Connective Tissue Growth Factor (CTGF) in Esophageal Squamous Cell Carcinoma
Qiang Han, Hua-Yong Zhang, Bei-Long Zhong, Xiao-Jing Wang, Bing Zhang, Hua Chen
Department of Thoracic and Cardiovascular Surgery, Fifth Affiliated Hospital of Sun Yat-Sen University, Zhuhai, Guangdong, China (mainland)
Med Sci Monit 2016; 22:3925-3934
This study investigated the mechanism of miR-145 in targeting connective tissue growth factor (CTGF), which affects the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of ESCC cells.
MATERIAL AND METHODS: A total of 50 ESCC tissues and their corresponding normal adjacent esophageal tissue samples were collected. Then, miR-145 expression in both ESCC clinical specimens and cell lines was detected using quantitative real-time PCR. CTGF protein was detected using immunohistochemistry. Dual luciferase reporter gene assay was employed to assess the effect of miR-145 on the 3’UTR luciferase activity of CTGF. Eca109 cells were transfected with miR-145 mimics and CTGF siRNA, respectively, and changes in cellular proliferation, migration, and invasion were detected via MTT assay, wound-healing assay, and Transwell assay, respectively. Western blotting assay was used to detect the expression of marker genes related to EMT.
RESULTS: MiR-145 was significantly down-regulated in ESCC tissues and cell lines compared with normal tissues and cell lines (P<0.05). We found significantly more positively expressed CTGF protein in ESCC tissues was than in normal adjacent esophageal tissues (P<0.01). Dual luciferase reporter gene assay showed that miR-145 can specifically bind with the 3’UTR of CTGF and significantly inhibit the luciferase activity by 55% (P<0.01). Up-regulation of miR-145 or down-regulation of CTGF can suppress the proliferation, migration, invasion, and EMT process of ESCC cells.
CONCLUSIONS: MiR-145 was significantly down-regulated in ESCC tissues and cell lines, while the protein expression of CTGF exhibited the opposite trend. MiR-145 inhibited the proliferation, migration, invasiveness, and the EMT process of ESCC cells through targeted regulation of CTGF expression.
Keywords: 3' Untranslated Regions, Carcinoma, Squamous Cell - pathology, Cell Line, Tumor, Cell Movement - genetics, Cell Proliferation - genetics, China, Connective Tissue Growth Factor - metabolism, Down-Regulation, Epithelial-Mesenchymal Transition - genetics, Esophageal Neoplasms - pathology, MicroRNAs - metabolism, Neoplasm Invasiveness, Transfection