端粒酶逆转录酶基因反义寡核苷酸抑制肺癌细胞端粒酶活性和诱导细胞凋亡

Inhibition of telomerase activity and induction of apoptosis in lung cancer cell by human telomerase reverse transcriptase gene antisense oligodeoxynucleotide

目的 观察端粒酶逆转录酶 (hTERT)基因反义寡核苷酸 (ASODN)对肺癌细胞端粒酶活性和细胞凋亡的影响。方法 实验分为ASODN组、正义寡核苷酸 (SODN)组和空白对照组 ,所用ASODN和SODN的浓度分别为 10 μmol/L ,脂质体为 16mg/L ,采用端粒重复扩增法、逆转录 聚合酶链反应、WesternBlot及流式细胞术分别观察各组端粒酶活性、hTERTmRNA和蛋白质表达以及细胞凋亡。结果 ASODN组显著下调或抑制肺癌细胞端粒酶活性和hTERT表达 ,但直到第 2 1天才出现细胞凋亡增多。

Objective To evaluate the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) on telomerase activity and apoptosis in lung cancer cell A549 line.Method Cultured cells were divided into three groups: ASODN(5′ GCGCGGCAGCGCGGGGGTGGCCG 3′), sense oligodeoxynucleotide(5′ CGGCCACCCCCGCGATGCCGCGC 3′,SODN) and control. The concentration of oligodeoxynucleotide and lipsome was 10 μmol/L and 16 mg/L respectively. The changes of telomerase activity, expression of hTERT mRNA and protein and apoptosis were observed in different groups. Results hTERT mRNA and protein levels, as well as telomerase activity began to be down regulated or inhibited when A549 cells were treated with ASODN for 72 h, but apoptosis was induced until cells were treated for 21 days with ASODN(rate of apoptosis was 0.88%, 2.77% and 12 00% for the control, SODN and ASODN group respectively). Conclusion It is suggested that hTERT ASODN might specially inhibit telomerase activity at translation level in lung cancer cells and it is further proved that the hTERT gene have significant correlation with telomerase activity. Further evidence is needed to prove whether hTERT ASODN might be a potential tool for the treatment of lung cancer.