hTERT基因反义核酸对Jurkat细胞端粒酶活性影响

Effect of antisense oligodeoxynucleotide of human telomerase reverse transcriptase on the telomerase activity of Jurkat cells

目的 :检测hTERT基因反义核酸对Jurkat细胞端粒酶活性的影响及其机制。方法 :TRAP法检测端粒酶活性 ,流式细胞仪检测hTERT蛋白表达 ,逆转录 -多聚酶链式反应检测hTERTmRNA表达。结果 :检测端粒酶活性 ,Jurkat细胞吸光度A值为 0 492± 0 0 5 1,hTERT反义核酸作用 48hA值降为 0 35 1± 0 0 5 1,hTERT反义核酸作用72hA值降为 0 2 38± 0 0 2 4;检测hTERT蛋白表达 ,Jurkat细胞hTERT蛋白阳性率 89 5 13%± 3 389% ,hTERT反义核酸作用 48hhTERT蛋白阳性率低至 77 2 37%± 2 872 % ,hTERT反义核酸作用 72hhTERT蛋白阳性率低至47 76 7%± 1 32 6 %。而hTERT正义核酸无上述作用。hTERT反义核酸作用Jurkat细胞 48h、72h对hTERTmRNA表达无影响。结论 :hTERT反义核酸降低Jurkat细胞端粒酶活性 ,机制可能是降低hTERT蛋白表达量 ,对hTERTmRNA无影响。

AIM: To observe the effect of antisense oligodeoxynucleotide of human telomerase reverse transcriptase(hTERT) on the telomerase activity in Jurkat cells and its mechanism. METHODS: The telomerase activity was detected by TRAP. hTERT protein and mRNA expressions were detected by flowcytometry and RT-PCR. RESULTS: The absorbency of the cells ( A values) was used to represent the telomerase activity. At 48 h and 72 h, the A values of the Jurkat cells treated with antisense oligodeoxynucleotide of hTERT decreased to 0.351±0.051 and 0.238±0.024, respectively, compared to the A value of 0.492±0.051 in the control cells without treatment ( P< 0.05). The positive rates of hTERT protein expression in Jurkat cells treated with antisense oligodeoxynucleotide of hTERT at 48h and 72h were 77.2%±2.9% and 47.8%±1 3%, respectively, compared to the positive rate of 89.5%±3.4% in the control cells without treatment ( P <0.05). No change of hTERT protein expression was observed when sense oligodeoxynucleotide of hTERT was used. However, antisense oligodeoxynucleotide of hTERT did not affect the expression of hTERT mRNA at 48 h and 72 h treatment. CONCLUSION: Antisense oligodeoxynucleotide of hTERT downregulated the telomerase activity and hTERT protein expression in Jurkat cells, but did not alter hTERT mRNA content.