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为了研究端粒酶催化亚基hTERT基因在癌变细胞中组成型表达的调控机制 ,采用凝胶阻滞电泳 (EMSA)实验方法检测人粒系白血病细胞HL 6 0、人红系白血病细胞K5 6 2、人肺癌细胞A5 4 9、人肝癌细胞HepG2及正常人肺成纤维细胞 2BS等体外传代的肿瘤和正常二倍体细胞核提取物中与hTERT启动子核心序列结合的核因子活性。结果在 4种实验肿瘤细胞中均可检测出与hTERT基因启动子结合的核因子活性 ,而正常二倍体细胞中则不存在这种活性的核因子。实验结果提示hTERT基因启动子结合因子可能是细胞癌变过程出现的特殊基因表达调控因子 ,与端粒酶在各种肿瘤细胞中广泛重新激活有关 ,有必要进一步分离鉴定这些肿瘤特异性的转录因子 ,研究其在细胞癌变及肿瘤细胞异常生物学表型形成中的作用
In order to study the regulatory mechanism of the expression of telomerase catalytic subunit hTERT gene in cancerous cells, gel electrophoresis electrophoresis (EMSA) assay was used to detect human granulocytic leukemia cell line HL 60 and human erythroid leukemia cell K5 6 2 . Human lung cancer cell A549, human liver cancer cell HepG2, and normal human lung fibroblasts 2BS and other in vitro passaged tumors and normal diploid nuclear extracts bind to the nuclear factor activity of the hTERT promoter core sequence. Results The activity of nuclear factor bound to hTERT gene promoter was detected in all four kinds of experimental tumor cells, but there was no such nuclear factor in normal diploid cells. The experimental results suggest that the hTERT gene promoter binding factor may be a special gene expression regulation factor in the process of cell carcinogenesis, and is related to the extensive reactivation of telomerase in various tumor cells. It is necessary to further isolate and identify these tumor-specific transcription factors. To study its role in cell carcinogenesis and abnormal biological phenotype of tumor cells