Effects of Exogenous CC10 Transfection on CyclinD1 Protein and mRNA Expression in A549 Lung Cancer Cells

摘要

Objective: To investigate the effects of exogenous CC10 gene transfection on cell cycle and the expression of cyclinD_1 protein and mRNA in A549 cells. Methods: A549 cells in all test groups (group A to E) and control group (group F) were transfected with exogenous CC10 gene by liposome for 8, 16, 24, 36, 48 and 0 h respectively. CC10 protein expression was detected in A549 cells by Western blot. The growth inhibitory rate was detected by MTT method. Flow cyometry analysis (FCS) and AnnexinV-PI staining were used to determine the changes of cell cycle progression and apoptosis rate in all groups. CyclinD_1 protein and mRNA expression in A549 cells was detected by the methods of immunocytochemistry and RT-PCR. Results: Exogenous CC10 gene could inhibit the growth of A549 cells, and the growth inhibitory rates in all test groups (from group A to E) were 24.7%, 33.1%, 44.3%, 61.7% and 74.2% respectively, and that in group F was 6.24%. CC10 blocked the cell cycle progression at G_0/G_1 and induced apoptosis gradually. In A549 cells of test groups, the expression of cyclinD_1 protein and mRNA was significantly decreased. Conclusion: The inhibitory effects of the transfection of exogenous CC10 gene on G_0/G_1 cycle of lung cancer cells might be related with the down-regulation of cyclinD_1 gene.