Effect of Na~+/dicarboxylate transporter protein on cell cycle and cyclin P16 in human fibroblasts / 中国组织工程研究

ABSTRACT

BACKGROUND: Na+/dicarboxylate transporter protein (NaDC3) participates in the metabolism of energy, which may has effect on cellular senescence; however, the mechanism is not clear. OBJECTIVE: To investigate the role of human NaDC3 in cellular senescence by means of cell cycle and P16 measurements. DESIGN, TIME AND SETTING: The single sample observation was performed in the Nanjing Drum Tower Hospital from December 2007 to May 2008. MATERIALS The WI-38 cell was purchased from ATCC Company, the retrovirus vector pLNCX2, sense vector PLNCX2-hNaDC3 and antisense vector PLNCX2-AshNaDC3 was provided by Clontech Company. METHODS: hNaDC3 was introduced into WI-38 cells with retroviral virus-mediated transfection. The integration and expression of exotic gene were confirmed. Whereafter, to examine the effects of hNaDC3 on cellular senescence of WI-38 cells by cell cycle and expression of P16. MAIN OUTCOME MEASURES: The integration and expression of hNaDC3, the cell morphology and generation of WI-38, the cell cycle and expression of P16 were observed. RESULTS: The results of Northern blot and Western blot revealed that hNaDC3 were expressed in WI-38/ hNaDC3 cells at gene and protein level, while, in WI-38/As hNaDC3 cells, there were integration of hNaDC3 mRNA with not protein expression. Compared with the control cells, WI-38 cells transfect with hNaDC3 cDNA showed cell generation decreased by eight to twelve passages, which in a low cell growth rate with cell cycle arrested at G1 phase, and the expression of P16 was elevated. CONCLUSION: Human NaDC3 gene may contribute to the process of cellular senescence of human diploid fibroblasts by increasing expression of P16.