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目的探讨下调线粒体转录因子A(mitochondrial transcription factor A,TFAM,也称为m TFA)对人表皮干细胞(epidermal stem cell,ESC)增殖、凋亡等的影响。方法将TFAM基因干扰慢病毒转染至ESC中,得到目标ESC为干扰组,并取对照病毒转染ESC作为对照组。对比分析两组中ESC的增殖与克隆成型率、凋亡以及端粒酶反转录酶表达水平等生物学特征的差异。结果两组比较,干扰组ESC克隆形成率及凋亡均显著高于对照组,而增殖率显著低于对照组,差异有统计学意义(P<0.05);在细胞端粒酶反转录酶水平测定中,两组均为阳性,但干扰组经定量分析,其平均光密度与阳性面积值均显著低于对照组,差异有统计学意义(P<0.05)。结论 TFAM基因的沉默可导致ESC体外增殖活性降低,促进其凋亡过程,通过增加克隆形成效率而利于维持干细胞特性,并可能进一步抑制创面的愈合。
Objective To investigate the effects of mitochondrial transcription factor A (TFAM), also known as m TFA, on the proliferation and apoptosis of human epidermal stem cells (ESC). Methods Transfection of lentivirus with TFAM gene into ESC resulted in the target ESC being the interference group and the control virus transfected with ESC as the control group. The biological characteristics of ESC, such as proliferation, cloning rate, apoptosis and expression of telomerase reverse transcriptase, were compared between the two groups. Results Compared with control group, the formation rate and apoptosis of ESC in interference group were significantly higher than those in control group, but the proliferation rate was significantly lower than that in control group (P <0.05). In telomerase reverse transcriptase In the level determination, both groups were positive, but the quantitative analysis of interference group, the average optical density and positive area values were significantly lower than the control group, the difference was statistically significant (P <0.05). Conclusion The silencing of TFAM gene leads to the decrease of proliferation and apoptosis of ESCs in vitro, and the maintenance of stem cell characteristics by increasing the efficiency of colony formation, which may further inhibit the wound healing.