端粒酶反义寡核苷酸对食管癌细胞Eca-109端粒酶活性及细胞凋亡的影响

目的探讨人端粒酶反义寡核苷酸(PS-ASODN)对食管癌Eca-109端粒酶活性及细胞凋亡的影响。方法1-5uM的PS-ASODN、5uM的N-ASODN作用于Eca-109细胞后,在倒置显微镜下连续观察Eca-109细胞形态学改变;采用半定量TRAP-银染法检测端粒酶活性;用流式细胞仪检测细胞凋亡率及细胞周期变化。结果PS-ASODN对Eca-109细胞具有生长抑制作用,PS-ASODN作用后,细胞的生长速度缓慢,细胞体积缩小,部分细胞变形漂起。随着药物浓度增加,PS-ASODN对Eca-109细胞端粒酶活性的抑制作用逐渐增强,呈浓度依赖性和序列特异性。流式细胞仪检测到凋亡峰并受阻于G0/G1期,而对照组寡核苷酸无上述作用。结论PS-ASODN不但抑制食管癌的增殖,降低端粒酶的活性,而且具有促凋亡的作用,对食管癌的临床治疗具有重要的价值。

Effect of telomerase antisense oligodeoxynucleotide ontelomerase activiyand apoptosis ofesophageal neoplasm

Objective To study the effect of telomerase antisense oligodeoxynucleotide(PS-ASODN)on telomerase activity and apoptosis of Eca-109 cell line.Methods Eca-109 cell line was transfected with 1-5uMPS-ASODN and 5UM N-ASODN.Microscope was devered tO examine the morphologic changes Of cell after treatment.Telomerase activity was detected by PCRsilver staining semi-quantitative TRAPassay.FlowCytometry was adopteded to examine apoptotic rate and cell cycle.Results PS-ASODN inhibited significantly the growth of Eca-109 cell line.Cells became round,suspended andsmall.The effect was in a dose dependent and sequence specific manner.The apoptotic peak was detected with FCM and cells were stayed in G0/G1 stage.Control oligodeoxynucleotide had no inhibiting effect on Eca-109 cell line.Conclusions PS-ASODN targeted hTR not only inhibited Eca-109 cellproliferation and telomerase activity,but also induced apoptosis.Thereforeit may be ane wtarget tO the esophageal neoplasm.