Construction and identification of lentiviral vector of micorRNA-206 gene

Objective To construct a recombinant lenticiral vector of microRNA-206 (miR-206) gene.Methods Oligo-chain DNA complemented to miR-206 gene was designed and lentivirus-based vectors was constructed.Those colonies with positive polymerase chain reaction (PCR) were sequenced.All virus stocks were transfected by Lipofectamine 2000.The titer of virus was tested according to the expression level of green fluorescent protein.The expression of miR-206 in MCF-7 infected by the lentivirus was analyzed via real-time reverse transcriptase-polymerase chain reaction (RT-qPCR).The proliferation of MCF-7 was tested by cell counting kit-8 (CCK-8).The changes of apoptosis and cell cycle was studied by flow cytometry.Results The lentivirus vector of miR-206 (LV-hsa-miR-206) were successfully constructed.After MCF-7 was infected by LV-hsa-miR-206,the expression of miR-206 gene was significantly increased from 1.000 to 120.331 ± 1.801.The proliferation of MCF-7 was decreased from 1.420 ±0.052 to 1.275 ±0.147 (2nd day),from 1.762 ±0.089 to 1.387 ±0.093 (3rd day),from 2.045 ±0.235 to 1.269 ±0.103 (4th day).The cell number of apoptosis and in G2 phase was increased.Conclusion The LV-hsa-miR-206 is construted successfully and may be used to infect MCF-7 cells.After MCF-7 cells infected by this sort of vectors,the experession of miR-206 and biological ability is significantly changed.LV-hsa-miR-206 provides the basis for research on miR-206 in breast cancer. Key words: MicroRNA-206 ;  Recombinant lenticiral vector;  Breast cancer;  Metastasis