hTERT基因永生化人毛乳头细胞系的转化特征

Transformed phenotype of immortalized human dermal papilla cell line induced by hTERT gene

目的探讨hTERT基因永生化人毛乳头细胞系是否具有转化特征。方法采用脂质体转染法,将hTERT基因导入体外培养的正常人毛乳头细胞,经G418筛选得到阳性克隆,连续传代培养。应用RT-PCR法检测hTERT mRNA的表达。采用细胞形态学观察、染色体核型分析、软琼脂克隆形成实验和裸鼠致瘤实验分析转染细胞是否具有转化特征。结果转染后获得1个阳性细胞克隆,扩大培养后细胞生长良好,现已连续传至50代。检测证实转染细胞稳定表达hTERT mRNA,细胞形态与未转染细胞基本相同,染色体核型正常,在软琼脂内不能生长,无裸鼠致瘤性。结论hTERT基因永生化人毛乳头细胞系基本上是正常细胞而无明显转化特征。

Objective To explore whether the immortalized human dermal papilla cell line induced by human telomerase reverse transcriptase（hTERT） gene has transformed phenotype. Methods The hTERT gene was transfected into cultured normal human dermal papilla cells, then the positive cells were selected with G418 and expanded for continuous culture. The expression of hTERT mRNA in the transfected cells was detected by RT-PCR. The morphology of the transfected cells was observed and the karyotype, growth in soft agar and tumorigenicity in nude mice of the transfected cells were analyzed. Results A positive cell clone has been selected,and the 50nd passage of transfected dermal papilla cells has been obtained so far. Expression of hTERT gene was detected on mRNA levels. The morphology of the transfected cells was similar to that of untransfected cells. The transfected cells exhibited a normal karyotype,were not able to grow in soft agar and were non-tumorigenic in nude mice. Conclusion The immortalized human dermal papilla cell line induced by hTERT gene is a normal cell line without obvious transformed phenotype.