Effect of InnVit gene expression on proliferation, apoptosis and cell cycle of mouse melanocyte cell line B10BR

Objective To assess the effect of InnVit gene expression on cell proliferation, apoptosis and cell cycle of B10BR cells, and to explore the biological function of lnnVit gene and its influence on the loss of melanoeytes in vitiligo. Methods InnVit gene-targeting siRNA was synthesized, and overexpression plasmid P3XF-P120 was constructed： BIOBR cells were classified into 4 groups to be transfected with control siRNA （negative control）, siRNA targeting InnVit gene （siRNA group）, empty plasmid （plasmid control group）, and P3XF-P120 plasmid （P3XF-P120 group）, respectively. After additional culture for various periods, the mRNA and protein levels of InnVit gene were detected by RT-PCR and Western blot, respectively, cell survival by MTF assay, cell cycle and apoptosis by flow cytometry. Results The expression of InnVit mRNA and protein significantly increased in P3XF-P120 group compared with the plasmid control group, but decreased in siRNA group compared with the negative control group （all P 〈 0.01）. After the transfection, the survival rate of cells was significantly lower in siRNA group than in negative control group at 24, 48 and 72 hours, but higher in P3XF-P120 group than in plasmid control group at 48 and 72 hours （all P 〈 0.01 ）. At 48 hours after the transfection, the early apoptosis rate and proportion of cells in GI phase increased from （10.24 ± 1.00）% and （56.11 ± 5.46）% in negative control group to （37.34 ± 3.26）% and （69.76 ± 6.08）%, in siRNA group, respectively （P〈 0.01 or 0.05）, from （14.58 ± 1.49）% and （55.14 ± 5.65）% in plasmid control group to （8.43 ± 0.86）% and （29.33 ± 3.01 ）%, in P3XF-P120 group, respectively （both P 〈 0.01 ）. Conclusions The suppression of InnVit gene efficiently inhibits the proliferation and induces apoptosis of B10BR cells, and vice versa. Therefore, the regulation of cell cycle may affect the proliferation of B10BR cells. Key words: InnVit gene;  Vitiligo;  Cell proliferation;  Apoptosis;  Cell cycle;  Cells, cultured