Jian-Xin Pang1, Xi-Yuan Chen, Shu-Guang Wu. 1. Institute of Pharmaceutic Sciences, First Military Medical University, Guangzhou 510515, China. pjx@fimmu.edu.cn
Abstract
OBJECTIVE: 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. METHODS: 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). RESULTS: 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. CONCLUSION: Transcription factor Sp1, but not Sp3, regulates telomerase activity by altering hTERT mRNA levels in Jurkat T cells.
OBJECTIVE: To investigate the role of transcription factors Sp1 and Sp3 in the regulation of telomerase activity and humantelomerase reverse transcriptase (hTERT) in Jurkat T cells. METHODS: By way of lipofectamine, Sp1 and Sp3expression 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). RESULTS: 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. CONCLUSION: Transcription factor Sp1, but not Sp3, regulates telomerase activity by altering hTERT mRNA levels in Jurkat T cells.
Authors: Lisandro Cunci; Marina Martinez Vargas; Roman Cunci; Ramon Gomez-Moreno; Ivan Perez; Abel Baerga-Ortiz; Carlos I Gonzalez; Carlos R Cabrera Journal: RSC Adv Date: 2014-10-15 Impact factor: 3.361