Inhibition of lung cancer Calu-6 cell line proliferation using Cdc42 gene shRNA
Volume 8, Issue 3, Winter 2018, Pages 261-270
https://doi.org/10.52547/JCT.8.3.261
Z Ghambari, i M Nabiun, H Jalali, L Karimzadeh
Abstract Aim: In current study, we aimed to reduce Cdc42 gene expression in lung carcinoma related cells, Calu-6, and assayed its effect on cell proliferation.
Material and Methods: To reduce the expression of Cdc42 gene shRNA system was used and lentiviral system was selected to deliver Cdc42 specific shRNA to Calu-6 cells. Recombinant lentiviruses produced by co-transfection of pMD2G, psPAX2 and p-GFP-C-shLenti plasmids into 293T cells using lipofectamin. Efficiency of transfection and transduction assessed by florescent microscopy. Viability of cells treated by recombinant lentiviruses assessed by MTT assay.
Results: florescent microscopy showed 80% transfection of 293T cells and high rate of Calu-6 cells transduction. MTT assay results revealed that viability of transduced Calu-6 cells reached to %58 and %40 in compare to control and negative control cells, respectively.
Conclusion: recombinant lentiviruses properly transfer Cdc42-shRNA into Calu-6 cells, leading to reduction of cell proliferation. Silencing of Cdc42 gene expression using lentiviruses is persist and long-term effect which can be under attention for gene therapy of lung cancer.