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Absence of telomerase activity and telomerase catalytic subunit mRNA in melanocyte cultures

机译:黑色素细胞培养中缺乏端粒酶活性和端粒酶催化亚基mRNA

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The classic model of activation of telomerase, for which activity has been found in most cancers including cutaneous malignant melanoma (CMM), dictates that enzyme activity is generated by pathological reactivation of telomerase in telomerase-negative somatic cells. However, recent data demonstrated physiological up-regulation in some normal cell types when established as proliferating cultures, indicating that, in some cancer types, telomerase is expressed by the process of up-regulation in telomerase-competent precursor cells. In this study, cultures of epidermal melanocytes, progenitor cells of CMM, were established and harvested in the logarithmic phase of growth. Telomerase activity was looked for using a non-isotopic variant of the telomeric repeat amplification protocol, and transcript expression of the hTERT gene, the rate-limiting catalytic telomerase subunit, was investigated by the reverse transcription polymerase chain reaction. Neither telomerase activity nor hTERT mRNA could be detected in proliferating melanocyte cultures. Our in vitro data argue against the model of telomerase as a common biomarker of cell proliferation. The results further suggest that telomerase is tightly controlled in normal melanocytes, and that telomerase is reactivated rather than up-regulated in melanocytic precursors during melanoma initiation or progression. ? 2000 Cancer Research Campaign
机译:端粒酶激活的经典模型已在包括皮肤恶性黑色素瘤(CMM)在内的大多数癌症中发现了其活性,该模型表明酶活性是由端粒酶阴性体细胞中端粒酶的病理性激活产生的。然而,最近的数据表明当建立为增殖培养物时,某些正常细胞类型中的生理上调,表明在某些癌症类型中,端粒酶通过端粒酶活性前体细胞中的上调过程表达。在这项研究中,建立了表皮黑素细胞(CMM的祖细胞)培养物,并在对数生长期收获。使用端粒重复扩增方案的非同位素变体寻找端粒酶活性,并通过逆转录聚合酶链反应研究hTERT基因(限速催化端粒酶亚基)的转录表达。在增殖的黑素细胞培养物中,端粒酶活性和hTERT mRNA均未检测到。我们的体外数据反对端粒酶作为细胞增殖的常见生物标志物的模型。结果进一步表明,端粒酶在正常黑素细胞中受到严格控制,端粒酶在黑素瘤发生或发展过程中在黑素细胞前体中被重新激活而不是上调。 ? 2000年癌症研究运动

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