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人端粒酶逆转录酶基因反义寡脱氧核苷酸对肺癌细胞端粒酶活性的抑制及凋亡诱导作用

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

作者信息

Wang Xiaoyang, Zhang Zhenxiang, Xu Yongjian, Chen Shixin, Xiong Weining

机构信息

Research Laboratory of Respiratory Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Zhonghua Nei Ke Za Zhi. 2002 Mar;41(3):175-8.

Abstract

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 micromol/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.

摘要

目的

评估人端粒酶逆转录酶(hTERT)基因反义寡脱氧核苷酸(ASODN)对肺癌细胞A549株端粒酶活性及细胞凋亡的影响。

方法

将培养的细胞分为三组:ASODN组(5'-GCGCGGCAGCGCGGGGGTGGCCG-3')、正义寡脱氧核苷酸组(5'-CGGCCACCCCCGCGATGCCGCGC-3',SODN)和对照组。寡脱氧核苷酸和脂质体的浓度分别为10 μmol/L和16 mg/L。观察不同组中端粒酶活性、hTERT mRNA和蛋白表达及细胞凋亡的变化。

结果

用ASODN处理A549细胞72小时后,hTERT mRNA和蛋白水平以及端粒酶活性开始下调或受到抑制,但直到用ASODN处理细胞21天才诱导细胞凋亡(对照组、SODN组和ASODN组的凋亡率分别为0.88%、2.77%和12.00%)。

结论

提示hTERT ASODN可能在翻译水平特异性抑制肺癌细胞中的端粒酶活性,进一步证明hTERT基因与端粒酶活性有显著相关性。是否hTERT ASODN可能成为治疗肺癌的潜在工具还需要进一步证据证实。

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