摘要
目的 探讨端粒酶反义寡聚脱氧核苷酸 (PS ASODN)对SGC 790 1胃癌细胞端粒酶活性及细胞凋亡的影响。方法 脂质体介导的PS ASODN作用于SGC 790 1细胞后 ,台盼蓝拒染法计算细胞生长抑制率 ,采用半定量TRAP 银染法检测端粒酶活性 ,用流式细胞仪观察细胞凋亡率及细胞周期变化 ,采用光镜及电镜观察细胞凋亡形态。结果 终浓度为 3、2及 1μM的PS ASODN对SGC 790 1细胞均有抑制作用 ,抑制率与对照组比较 ,差异有显著性 (P <0 .0 1) ;PS ASODN转染后 48h ,端粒酶活性明显降低 (P <0 .0 5 )。以上抑制作用呈剂量依赖性及序列特异性。流式细胞仪检测到凋亡峰 ,细胞受阻于G0 /G1期 ;光镜及电镜显示典型的细胞凋亡形态改变。对照寡核苷酸无上述作用。结论 以端粒酶RNA模板区为靶点的反义寡核苷酸不但明显抑制胃癌细胞的增殖 ,降低端粒酶活性 ,而且具有促凋亡作用 ,对胃癌具有重要治疗价值。
Objective To study the effects of telomerase antisense oligodeoxynucleotides (PS ASODN) on telomerase activity and apoptosis of gastric carcinoma cell line.Methods SGC 7901 cell line was transfected with PS ASODN mediated by liposome (DOTAP). The inhibition rate was determined by trypan blue staining. Semi quantity TRAP assay was used to detect telomerase activity. Flow cytometry was adopted to examine apoptotic rate and cell cycle. Cell morphologic changes were examined under light and electronic microscope.Results 1 3?μM PS ASODN inhibited significantly the growth of SGC 7901 cell line and the inhibition rate was higher than that in control group( P <0.01). PS ASODN reduced telomerase activity( P <0.05) from 48?h after transfection. The effect was in dose dependent and sequence specific manner. The apoptotic peak was detected with FCM and cells were stayed in G 0/G 1 stage. Typical apoptotic morphologic feature was discovered under electronic and light microscope. Control oligodeoxynucleotide had no inhibiting effect on SGC 7901 cell line.Conclusions PS ASODN targeted hTR not only inhibited SGC 7901 cell proliferation and telomerase activity, but also induced apoptosis. Therefore it may be a new target to gastric carcinoma therapy.
出处
《临床内科杂志》
CAS
北大核心
2002年第4期278-280,T001,共4页
Journal of Clinical Internal Medicine
