Construction of recombinant lentivirus vector containing retinoic acid-induced gene G and its effect on human lung cancer A549 cell line / 中国癌症杂志

ABSTRACT

Background and purpose:Retinoic acid-induced gene G (RIG-G) is a tumor suppressor gene which is cloned by NB4 cell line from a acute promyelocytic leukemia cell. This study aimed to investigate the effect ofRIG-G in lung cancer cells A549 by constructing a lentiviral vector expressing RIG-G under doxycycline (DOX) regulation.Methods:RIG-G gene ampliifcation was performed by quantitative real-time PCR (qRT-PCR). pLenti6/TO/V5-GIM-RIG-G lentiviral vector withGFP was built by LR recombination system. The concentration of pLenti6/TO/V5-GIM-RIG-G lentiviral vector andTet-on lentiviral vector were measured by virus titer method. After infecting A549 cells, stably transfected lines were selected via limiting dilution analysis.RIG-G gene expression was examined by immunolfuorescence staining and Western blot assay. Cellular proliferation was determined by CCK-8 assay.Results:The concentrations of pLenti6/TO/V5-GIM-RIG-G lentiviral vector andTet-on lentiviral vector were 1.0×108TU/mL and 4×109 VP/mL, respectively. RIG-G was expressed in lentivirus infected A549 cells after adding DOX, and the amount of cells withGFP could be observed by lfuorescence microscopy.After the expression of RIG-G protein, the prolif-eration activity of A594 cell was signiifcantly inhibited compared to the control group (1.168±0.107vs 2.099±0.162, P<0.05).Conclusion:The regulated expression ofRIG-G gene was established in A549 lung cancer cell line. The RIG-G protein has potential abilities to inhibit the proliferation of lung cancer cell A549.