hTERT基因反义核酸对A2780细胞端粒酶活性影响

Effect of antisense oligodeoxynucleotide of human telomerase reverse transcriptase on the telomerase activity of A2780 cells

目的:检测hTERT基因反义核酸对A2780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。结果:检测端粒酶活性,A2780细胞吸光度A值为(0.492±0.051),hTERT反义核酸作用48 h A值降为(0.351±0.051),hTERT反义核酸作用72 h A值降为(0.238±0.024)检测hTERT蛋白表达,A2780细胞hTERT蛋白阳性率(89.513±3.389)%,hTERT反义核酸作用48hhTERT蛋白阳性率低至77.237±2.872%,hTERT反义核酸作用72 h hTERT蛋白阳性率低至(47.767±1.226)%。而hTERT正义核酸无上述作用。hTERT反义核酸作用A2780细胞48 h、72h对hTERTmRNA表达无影响。结论:hTERT反义核酸降低A2780细胞端粒酶活性,机制可能是降低hTERT蛋白表达量。

Objective： To observe the effect of antisense oligodeoxynucleotide of human tdomerase reverse transcriptase （hTERT） on the telomerase activity in A2780 cells and its mechanism. Methods： The telomerase activity was detected by TRAP. hTERT protein expressions were detected by flowcytometry. Results： The absorbency of the cells （ A values） was used to represent the telomerase activity. At 48 and 72 h, the A values of the A2780 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 A2780 cells treated with antisense oligodeoxynucleotide of hTERT at 48 and 72 h were 77. 237%±2. 872% and 47. 767%±1.226%, respectively, compared to the positive rate of 89. 513%±3. 389% in the control cells without treatment （P〈0. 05） . No change of hTERT protein expression was observed when sense oligodeoxynucleotide of hTERT was used. Conclusion： Antisense oligodeoxynucleotide of hTERT could downregulate the telomerase activity and hTERT protein expression in A2780 cells.