Construction of miR-373-3p lentiviral vector with low expression and its effect on proliferation and migration of endometrial cancer cell line HEC-1A

LI Meiyi; XU Hong; TAN Feifei

doi:10.3969/j.issn.1002-266X.2017.45.006

Abstract：Objective To construct the lentiviral vector of human miR-373-3p with low expression and to explore its effects on the proliferation and migration of endometrial carcinoma cell line HEC-1A. Methods PCR was used to amplify the target gene containing pre-miR-373-3p sequence,and we cloned it into GV280 vector with restriction enzymes to con-struct the GV280-pre-miR-373-3p inhibitor expression plasmid which was co-transfected with packaging plasmid into 293T cells. After viral packaging,the titer of virus was detected. Empty lentivirus (empty virus group)and recombinant lentivir-us harboring miR-373-3p (miR-373-3p group)were employed to infect human HEC-1A cells,respectively,and the control group was set up. RT-PCR was used to determine the expression of miR-373-3p in HEC-1A cells. The cell proliferation was measured by CCK-8 assay at 24,48,72,and 96 h after culture,and the cell migration was detected by Scratch test and Transwell assay at 24 and 48 h. Results Human miR-373-3p entiviral vector with lower expression was constructed suc-cessfully. Compared with the empty virus group and control group,the expression of miR-373-3p in the HEC-1A cells de-creased,and cell migration decreased at 48,72,and 96 h in the miR- 73-3p group (all P < 0. 05);HEC-1A cell scratch area mobility decreased,and the number of transmembrane cells decreased at 24 and 48 h (all P < 0. 05). No significant difference was found in the above parameters between the empty virus group and the control group (all P < 0. 05). Conclu-sion The lentiviral vector containing miR-373-3p with low expression is successfully constructed,and low expression of miR-373-3p inhibits the cell proliferation and migration of endometrial cancer cells HEC-1A.