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首页> 外文期刊>International medical journal: IMJ >Detection of TERT Amplification in Cancer Cells Using Fluorescence in situ Hybridization and Quantitative Real Time PCM Assays
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Detection of TERT Amplification in Cancer Cells Using Fluorescence in situ Hybridization and Quantitative Real Time PCM Assays

机译：荧光原位杂交和定量实时PCM分析检测癌细胞中的TERT扩增

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Background: High expression or activity of telomerase in malignant cells has been associated with copy number gain or amplification of TERT gene.Methodology: Fluorescence in situ hybridization (FISH) and quantitative real time-PCR (qPCR) assays were employed to detect TERT gene amplification status in cancer cell lines K562 and MDA-MB 157 in comparison with normal cells.Results: Both analyses produced concordant results in determining the TERT gene amplification of low passage number in K562 and MDA-MB 157 cells. However, no amplification of TERT was detected in high passage number K562 cells and normal cells using both methods. High passage number K562 cells demonstrated trisomy of chromosome 5 in FISH analysis whereas no TERT amplification was detected in qPCR which showed N value within normal range.Discussion and Conclusion: Other previous studies had demonstrated alterations in cell morphology, karyotype pattern, response to stimuli, growth rates and protein expression, in cell lines at high passage number compared to cells in lower passage. In the present study, both qPCR and FISH assays were successfully optimized in detection of TERT copy number status in K562, MDA-MB 157 and normal cells. The findings observed in both assays were concordant which indicated increased TERT copy number status in K562 and MDA-MB 157 cells. TERT amplification was detected in K562 and MDA-MB 157 cells with low passage number but not in cells with high passage number. From our results, it is reasonable to suggest that the qPCR assay is rapid and useful to be applied as a potential reliable diagnostic tool in clinical samples.
机译：背景：端粒酶在恶性细胞中的高表达或活性与TERT基因的拷贝数增加或扩增有关。方法：采用荧光原位杂交（FISH）和定量实时PCR（qPCR）方法检测TERT基因扩增结果：两种分析在确定K562和MDA-MB 157细胞中低传代数的TERT基因扩增方面产生了一致的结果。但是，使用这两种方法在高传代数的K562细胞和正常细胞中均未检测到TERT扩增。高传代数目的K562细胞在FISH分析中显示出5号染色体三体性，而qPCR中未检测到TERT扩增，其N值在正常范围内。讨论和结论：其他先前的研究表明，细胞形态，核型模式，对刺激的反应，与传代数较低的细胞相比，传代数较高的细胞系中的生长速率和蛋白质表达高。在本研究中，成功​​地优化了qPCR和FISH分析以检测K562，MDA-MB 157和正常细胞中TERT的拷贝数状态。在两种测定中观察到的发现是一致的，这表明在K562和MDA-MB 157细胞中TERT拷贝数状态增加。在低传代数的K562和MDA-MB 157细胞中检测到TERT扩增，而在高传代数的细胞中未检测到TERT扩增。根据我们的结果，可以合理地认为qPCR测定是快速有效的，可作为临床样品中潜在的可靠诊断工具使用。

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