Sp1对HL-60细胞人端粒酶逆转录酶表达的调控

Regulation of human telomerase reverse transcriptase expression in HL-60 cell by Sp1

目的:探讨Sp1转录因子在人白血病细胞株HL-60中对人端粒酶逆转录酶(hTERT)基因表达的影响。方法:电泳迁移率分析(EMSA)hTERT启动子区(-116～-88 bp)是否存在Sp1结合位点。RT-PCR检测经普卡霉素处理的细胞hTERTmRNA表达。结果:hTERT启动子序列(-116～-88 bp)与核蛋白作用,出现DNA-核蛋白结合条带,普卡霉素抑制Sp1与此序列结合。普卡霉素浓度依赖性下调hTERT mRNA表达,200 nmol/L组和100 nmol/L组均明显低于50 nmol/L组、25 nmol/L组和对照组(P<0.01)。结论:Sp1与hTERT启动子区(-116～-88 bp)结合,转录激活hTERT表达。

Objective ：To explore the regulation of human telomerase reverse transcriptase（hTERT） expression in HL-60 cell by Spl. Methods ：The Spl binding sites of hTERT promoter sequence（ -116 - -88 bp） were assayed by eleetrophoretic mobility shift assay （EMSA）. The hTERT mRNA expressions of HL-60 cells treated with different concentration of mithramycin were examined by RT-PCR. Results：The binding bands of DNA-protein were detected by autoradiography when nuclear protein interacted with hTERT promoter sequence. Mithramyein inhibited the binding of Spl and the sequences, and decreased the mRNA expression of hTERT with concentration dependent manner, and which of mithramyein of 200 nmol/L group and 100 nmol/L group were significantly lower than those of 50 nmol/L group ,25 nmol/L group and the control group（ P 〈 0.01 ）. Conclusions： Spl can bind the promoter sequence （ - 116 - -88 bp） ,which can upregulate the mRNA expression of hTERT.