端粒酶催化亚基反义寡核苷酸对食管癌细胞的抑制作用

Inhibition of antisense oligodeoxynucleotides of human telomerase reverse transcription on esophageal carcinoma

目的：探讨人端粒酶催化亚基（human telomerase reserse transcription，hTERT）反义寡核苷酸（antisense oligodeoxynucleotide，ASODN）对食管癌细胞株Eca-109端粒醇活性的影响，及其对癌细胞的生长抑制作用。方法：MTT比色法测定ASODN对Eca-109细胞的生长抑制率；倒置显微镜观察5μmol／L的ASODN作用于Eca-109癌细胞10d后的细胞形态学改变；端粒酶活性检测采用半定量TRAP-银染法。结果：ASODN对Eca-109细胞的生长抑制作用具有剂量及时间依赖性和序列特异性。ASDON作用后，Eca-109细胞的生长速度缓慢，细胞间连接疏松。细胞变圆漂起，细胞体积缩小。随作用时间延长，ASODN对Eca-109细胞端粒酶活性的抑制作用逐渐增强，1～15d，相对端粒酶活性95～22。呈时间-依赖性；随药物浓度增加，ASODN对Eca-109细胞端粒醇活性的抑制作用逐渐增强，1～5μmol／L相对端粒酶活性86～14。呈浓度-依赖性。结论：端粒酶催化亚基反义寡核苷酸对食管癌具有重要治疗价值，有望成为食管癌治疗的潜在途径。

OBJECTIVE： To evaluate the effect of human telomerase reverse transcriptase （hTERT） gene antisense oligodeoxynucleotide （ASODN） on telomerase activity of human esophagus cancer cell line Eca-109. METHODS： MTT clorimetry was used to determine the cell growth inhibitory rate. Microscope was adopted to examine the morphologic changes of cells after Eca-109 was given 5μmol/L PS-ASODN for 10 d. Telomerase activity was detected by silver staining semi-quantitative TRAP assay. RESULTS： ASODN could, inhibit telomerase activity and proliferation of Eca-109 cell line in a dose and period dependent manner retative telamerase activity： 1 -15 d, 95 - 22; 1 - 5 μmol/L 86- 14. The growth rate of cells was reduced by ASDON. Cells became round, suspended and small. CONCLUSION： ASODN of human telomerase reserse transcription can inhibit telomerase activity and cell proliferation of Eca-109 cell line, suggesting that ASODN implied an important value for treatment of esophagus cancer.