shRNA mediated inhibition of Cdc42 gene expression in Calu-6, lung cancer cells

Background InformationRNAi technique as a new strategy in gene therapy is the effective gene silencing method. Cdc42 is a member of Rho GTPases involving in lung cancer cells migration and proliferation. In the present study, targeting and inhibiting the Cdc42 expression in Calu-6 cells was investigated. Recombinant lentiviral particles were produced by co-transfection of pMD2.G, psPAX2 and pGFP-C-shLenti plasmid in 293T packaging cells. Calu-6 cells were transduced by recombinant lentiviruses using polybrene. GFP-fluorescence microscopy and MTT assay were used to assess the Calu-6 target cell transduction and rate of lentiviral transduced cells proliferation, respectively. Real time PCR was performed to compare the expression of Cdc42 gene before and after shRNA delivery.ResultsGFP-fluorescence microscopy analysis showed that Calu-6 cells were successfully transduced with recombinant lentiviral expressing shRNA-Cdc42. The viability of transduced cells was reduced within 72, 96 and 120 hours of transduction process. Real time PCR analyze showed the significant reduction of Cdc42 gene expression.Conclusions:Lentiviral vectors may be reasonable tools for this gene delivery due to stable expression of silencing RNA. Inhibition of Cdc42 expression by lentiviral mediated shRNA delivery could be an effective method to inhibit proliferation of lung cancer cells.Significance:Over expression of Cdc42 gene have been seen in lung cancer. This make Cdc42 gene a key target in treatment of cancer. On the other hand, gene therapy is a proper method to modulate gene expression. It seems modulation of cdc42 gene expression by gene therapy accompany with proper vehicles creating hopes in treatment of lung cancer.