Effect of antisense oligodeoxynucleotide on cell proliferation and telomerase activity in K562 cell line / 白血病·淋巴瘤

ABSTRACT

Objective To observe anti-sense phosphorothioate oligonucletide (ASPSODN) targeted directly to hTERT mRNA to its inhibiting effect on aimed gene and the influence on the telomerase activity, cellular proliferation, cell apoptosis of K562 cells. Methods Human leukemia cell line K562 was transfected with anti-sense oligonucleotide ASPSODN by liposome. The proliferation activity of K562 cell line was determined by using methyl thiazolyl tetrazolium assay, and telomerase activity was detected by TRAP-PCR-ELISA. Flow cytometry was adopted to examine apoptotic rate and cell cycle. RT-PCR was used to detect the expression of target gene hTERT mRNA. Results 0.6 μmol/L ASPSODN (0.42 ±0.16) was remarkably decreased the expression of hTERT mRNA, Telomerase relative activation was decreased by 52 %. According to 0.6 (μmol/L ASPSODN caused significant the inhibition of K562 cell growth. Apoptotic rate and cell cycle was examined by 0.6 μmol/L ASPSODN with flow cytometry. The cell apoptosis rate of 0.6μmol/L ASPSODN were 10.31 %. It showed the cells treated with 0.6 uU PSASODN arrested in G_1/C_0. The ratio of cells in G_2/M and S period was reduced. But there was no characteristic apoptosis peak. Conclusion ASPSODN targeted hTERT can inhibit the expression of target gene hTERT mRNA, and decrease the telomerase activity of K562 cells. ASPSODN can inhibit strongly the proliferation of K562 cell and induce cell apoptosis by decreasing telomerase activity.