The effect of recombinant lentiviral vector encoding miR-145 on human esophageal cancer cells

• Published in: Tumor biology Vol. 36; no. 12; pp. 9733 - 9738
• Main Authors: Wang, Tian-Yun, Zhang, Qing-qing, Zhang, Xi, Sun, Qiu-Li, Zhao, Chun-Peng, Wang, Xiao-Yin
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.12.2015; Springer Nature B.V
• Subjects: Apoptosis - genetics; Biomedical and Life Sciences; Biomedicine; Cancer; Cancer Research; Carcinoma - genetics; Carcinoma - pathology; Cell Cycle - genetics; Cell Line, Tumor; Cell Proliferation - genetics; Cells; Esophageal Neoplasms - genetics; Esophageal Neoplasms - pathology; Genetic Vectors; Green Fluorescent Proteins - genetics; Humans; Lentivirus - genetics; MicroRNAs - genetics; Molecules; Research Article; Ribonucleic acid; RNA; Esophageal carcinoma; MicroRNA; Metastasis; Lentiviralvector; Apoptosis
• ISSN: 1010-4283; 1423-0380
• DOI: 10.1007/s13277-015-3743-1

miR-145, a newly identified microRNA molecule, is hypothesized to function as a tumor suppressor, but this activity has not been investigated in esophageal l carcinoma (EC). The aim of this study was to investigate the effect of miR-145 on the biological features of EC cells. miR-145 was obtained using PCR technology and cloned into the lentiviral vector, pLVX-IRES-ZsGreen1, to construct the resulting vector, pLVX-IZ-miR-145. The vector was packaged, the viral titer was tested, and ECA109 cells were infected with the optimal viral titer. Cells that were stably transfected with miR-145 were screened. Flow cytometry was used to analyze enhanced green fluorescence protein gene expression, and to measure cell apoptosis and cell cycle. miR-145 expression was detected by real-time fluorescent quantitative PCR. Furthermore, cell proliferation was assayed using CCK-8 assay. The pLVX-IZ-miR-145 vector was successfully constructed, and the viral titer achieved up to 5.0 × 10 8 TU/mL. The transfection efficiency was 90 %. Compared to the control group, the expression level of miR-145 in the transfected group was significantly higher (185-fold, P  < 0.05). miR-145 overexpression significantly inhibited esophageal cancer cell proliferation ( P  < 0.05). Moreover, the number of cells at the G2/M stage, as well as the cell apoptotic rate, in the miR-145-transfected group was significantly increased ( P  < 0.05). Our study reveals that overexpression of miR-145 inhibits cell proliferation, increases apoptosis, and influences the cell cycle progression of EC cell.