hTERT激活人成牙本质细胞端粒酶的实验研究

Experimental study of hTERT activating telomerase in human odontoblast

目的 :明确外源性人端粒酶逆转录酶 (humantelomerasereversetranscriptase,hTERT)能否调控人成牙本质细胞的端粒酶活性。方法 :培养人成牙本质细胞样细胞 ,采用脂质体转染法 ,将编码hTERT基因的真核表达质粒pCIneo -hTERT导入目的细胞。培养液加G418筛选 ,稳定后检测hTERT在mRNA和蛋白水平的表达以及端粒酶活性。结果 :转染后细胞表达hTERTmRNA及其蛋白 ,同时端粒酶活性转为阳性。结论 :在人成牙本质细胞样细胞内 ,外源性hTERT可以激活端粒酶 ,为建立永生化细胞系奠定了基础。

AIM: To explicate whether the telomerase activity is regulated by human telomerase reverse transcriptase(hTERT)in human odontoblast. METHODS: Human odontoblast-like cells were cultured and transfected with eukaryotic expressing plasmid pCIneo-hTERT encoding hTERT . After selection with G418 to stablize the transfection, expression of hTERT mRNA and protein , and telomerase activity in untransfected and transfected cells were cultured. RESULTS: After stable transfection, hTERT was expressed at mRNA and protein level, and meanwhile telomerase activity was positive in these transfected cells. CONCLUSION: In human odontoblast-like cells, telomerase is activated by exogenous hTERT. This is the first step to establish immortalized human odontoblast cell line.