Pang Jian-Xin, Chen Xi-Yuan, Wu Shu-Guang
Institute of Pharmaceutic Sciences, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2002 Jun;22(6):481-5.
To investigate the role of transcription factors Sp1 and Sp3 in the regulation of telomerase activity and human telomerase reverse transcriptase (hTERT) in Jurkat T cells.
By way of lipofectamine, Sp1 and Sp3 expression vectors were transferred into Jurkat T cells in which the protein expression levels of Sp1 and Sp3 were examined by Western blotting. Telomerase PCR-ELISA was used to assess the telomerase activity, and telomerase-mediated DNA- ladders were analyzed by silver staining and hTERT mRNA level determined by reverse transcriptase-PCR (RT-PCR).
Treatment of Jurkat T cells with Sp1 expression vector for 36 h resulted in significant increase in Sp1 and Sp3 protein levels by 59.6% (P<0.01) and 36.8% (P<0.05) respectively. Enhancement of Sp1 expression obviously increased telomerase activity and hTERT mRNA levels at a rate of 38.5% (P<0.05) and 25.4% (P<0.05) respectively, whereas Sp3 evinced no significant effect on both telomerase activity and hTERT mRNA levels.
Transcription factor Sp1, but not Sp3, regulates telomerase activity by altering hTERT mRNA levels in Jurkat T cells.
研究转录因子Sp1和Sp3在调节Jurkat T细胞中端粒酶活性及人端粒酶逆转录酶（hTERT）中的作用。
通过脂质体转染法将Sp1和Sp3表达载体导入Jurkat T细胞，采用蛋白质免疫印迹法检测Sp1和Sp3的蛋白表达水平。采用端粒酶PCR-ELISA法评估端粒酶活性，通过银染分析端粒酶介导的DNA梯带，并采用逆转录聚合酶链反应（RT-PCR）测定hTERT mRNA水平。
用Sp1表达载体处理Jurkat T细胞36小时后，Sp1和Sp3蛋白水平分别显著升高59.6%（P<0.01）和36.8%（P<0.05）。Sp1表达增强明显提高了端粒酶活性和hTERT mRNA水平，分别提高了38.5%（P<0.05）和25.4%（P<0.05），而Sp3对端粒酶活性和hTERT mRNA水平均无显著影响。
转录因子Sp1而非Sp3通过改变Jurkat T细胞中hTERT mRNA水平来调节端粒酶活性。
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