全硫代反义寡核苷酸对肺癌细胞生长及端粒酶活性的影响

Effects of the Proliferation and Telomerase Activation on Human Lung Cancer Cell by Antisense Phosphorothioate Oligonucleotide

目的将互补于hTR模板区的全硫代修饰型反义寡核苷酸（PS-ASODN）作用于肺癌细胞NCI-H292，探讨其对肺癌的治疗价值。方法PS-ASODN经lipotap脂质体转染作用于NCI-H292细胞，采用MTF法、ELISA法和流式细胞术检测NCI-H292细胞的体外增殖、端粒酶活性、细胞凋亡和细胞周期的改变。结果①MTT法检测，1．0μmol／LPS-ASODN处理后24h细胞生长抑制率为12．86％，且随着时间的延长，抑制作用逐渐增强。②ELISA法检测结果，0．5～1．5μmoL／L的PS-ASODN对NCI-H292细胞作用72h后端粒酶皆为阴性，对照组阳性。③流式细胞仪检测结果：端粒酶PS-ASODN转染NCI-H292细胞培养72h后，检测到细胞早期凋亡率增加，凋亡率为15．3％，与对照组相比有极显著意义（P〈0．01）。PI染色结果细胞被阻止在G1／G0期，S期比率有所降低，降为14．5％。结论①端粒酶PS-ASODN对NCI-H292细胞增殖有明显的抑制作用，其抑制作用具有浓度、时间依赖性。②端粒酶PS-ASODN能明显地抑制NCI-H292的端粒酶活性。

Objective The aim is to investigate the treatment effect of phosphorothioate antisense oligonucleotide （PS-ASODN） on human lung cancer cell line NCI-H292. Methods Human lung cancer cell line NCI-H292 treated with PS-ASODN liposome. The anti-proliferation effect on lung cancer cells were determined by MTT assay. The activity of telomerase was tested by the way of ELSIA. Apoptosis and cell cycle were examined by flow cytometer method. Results ①NCI-H292 cells was inhibited by 1.0μmol/L telomerase PS-ASODN,the inhibitory rate was 12.86% after 24 h later. And with times gone. ②Telomerase of NCI-H292 cells was repressed by 0.5-1.5μmol/L telomerase PS-ASODN after 72 h later, it showed that telomerase PS-ASODN could inhibit the activity of telomerase. ③There were earlier spike when the NCI-H292 cells were treated with 1.5μmol/LPS-ASODN at 72 h were15.3% （ P 〈 0.01 ）, the apoptosis rate. It showed that cells treated with 1.5μmol/L PS-ASODN arrested in G1/G0,the ratio of cells in Speriod was reduced to 14.5%. Conclusion ①PS-ASODN had significant inhibitory effects on the proliferation of the NCI-H292 cells. The inhibitory effects were sequence specificity and density,time-depended.②PS-ASODN had significant inhibitory effects on the telomerase enzyme activity.