人高亲和力钠依赖二羧酸转运蛋白3对人成纤维细胞衰老进程的影响

Effect of human Na^+/dicarboxylate cotransporter 3 on human fibroblasts (WI-38) on the aging process

目的 探讨人高亲和力钠依赖二羧酸转运蛋白 3(hNaDC3)在人胚肺二倍体成纤维细胞 (WI 38)衰老中的作用。 方法 通过构建逆转录病毒载体 ,使用含有hNaDC3基因的逆转录病毒液将hNaDC3基因导入WI 38中 ,并获得表达 ,观察其对WI 38细胞衰老的影响。 结果 与对照细胞相比 ,hNaDC3基因导入后 ,WI 38细胞传代数减少 10～ 13代 ,生长速率降低 4 0 % ,细胞周期阻滞于G1期 ,细胞形态呈衰老细胞样变化 ,衰老相关 β 半乳糖苷酶 (SA β gal)染色阳性率上升 ,线粒体膜电位下降。 结论 hNaDC3可能促进人W 1 38衰老。

Objective To investigate the role of human Na +/dicarboxylate cotransporter 3 (hNaDC3) in replicative senescence of normal human embryonic lung diploid fibroblasts(WI 38). Methods hNaDC3 gene was introduced into WI 38 cells through infection with reconstructed retroviral virus vector and the expression of hNaDC3 was detected. Then the effects of hNaDC3 on replicative cellular senescence of WI 38 cells were examined. Results Compared with the control cells, WI 38 cells transfected with hNaDC3 cDNA showed significant suppression of growth rate by 40%, the cell cycle was stopped at G 1 phase. It also showed that the positive rate of SA β gal staining was increased and mitochondrial membrane potential was decreased. The morphological characteristics of senescent fibroblasts appeared during earlier passages. Conclusions hNaDC3 gene may contribute to the process of cellular senescence of human diploid fibroblasts.