Abstract:ObjectiveTo investigate the effects of RING finger and WD repeat domain(RFWD3)on cervical cancer cells proliferation and migration by constructing the recombinant lentivirus vector which carries RFWD3.MethodsThe gene fragment of RFWD3 protein was amplified with the cDNA of HeLa cells,and was cloned into the lentiviral vector pCDH-CMV-MCS-EF1-copGFP (pCDH) plasmid by DNA recombination technique to construct the recombinant eukaryotic expression-vector of pCDH-RFWD3.The plasmid was verified to be corrected by enzymatic digestion and sequencing,and then RFWD3 fusion virus was obtained by lentivirus packaging method,and infected human embryonic kidney epithelial cells (293T) by gradient dilution method to detect the virus titer.Western blot assay was used to detect the expression of RFWD3 protein in HeLa cells after RFWD3 fusion virus infection,and the effect of up-regulated RFWD3 expression on the proliferation and migration ability of HeLa cells was studied by CCK-8 method and cell scratch test.ResultsThe pCDH-RFWD3 plasmid carrying RFWD3 gene was successfully constructed.Three plasmids were co-transfected into Human Embryonic Renal Epithelial cells (293T) by lentivirus packaging to construct RFWD3 gene fusion lentivirus.The level of RFWD3 protein was significantly increased and the cell proliferation and migration activity were significantly increased in cervical cancer cells which infected with lentivirus(P<0.05).ConclusionStudies have demonstrated that RFWD3 can significantly promote the proliferation and migration ability of HeLa cells.