Bièche I, Noguès C, Paradis V, Olivi M, Bedossa P, Lidereau R, Vidaud M
Laboratoire de Génétique Moléculaire, Faculté des Sciences Pharmaceutiques et Biologiques de Paris, France.
Clin Cancer Res. 2000 Feb;6(2):452-9.
Recent observations support the notion that telomerase expression is essential for the formation of human tumor cells [W-C. Hahn et al., Nature (Lond.), 400: 464-468, 1999]. The expression pattern of hTERT, the human telomerase catalytic subunit gene, is a rate-limiting determinant of the enzymatic activity of human telomerase. We have developed a real-time quantitative RT-PCR assay based on Taq-Man fluorescence methodology to quantify the full range of hTERT mRNA copy numbers. We validated the method on a series of 134 unilateral invasive primary breast cancer patients with known long-term outcome. Three-quarters of the breast tumors (75.4%; 101 of 134) were hTERT positive, i.e., contained detectable and quantifiable hTERT mRNA. hTERT-positive patients had significantly shorter relapse-free survival (P = 0.017) after surgery compared with hTERT-negative patients. The prognostic significance of hTERT status persisted in Cox multivariate regression analysis. When we subdivided hTERT-positive patients (n = 101) into three equal groups (tumors showing small, intermediate, or high increase in hTERT mRNA content), we observed statistical (or a trend toward) links between high hTERT mRNA levels and Scarff-Bloom-Richardson histopathological grade III (P = 0.066), and negative estrogen (P = 0.002) and progesterone (P = 0.048) receptor status, and therefore with higher aggressiveness of breast tumors. High hTERT mRNA levels were also linked to MYC gene overexpression (P = 0.007). These findings show that the quantitative evaluation of hTERT mRNA can have important prognostic significance in human breast cancer. In addition, our simple, rapid, and semiautomated assay method is suitable for routine hTERT mRNA detection and quantification and will be a powerful tool in large, randomized, prospective, cooperative group trials and in the hTERT-based therapy project.
最近的观察结果支持这样一种观点，即端粒酶表达对于人类肿瘤细胞的形成至关重要[W-C. 哈恩等人，《自然》（伦敦），400: 464 - 468，1999]。人端粒酶催化亚基基因hTERT的表达模式是人类端粒酶酶活性的限速决定因素。我们基于Taq-Man荧光方法开发了一种实时定量RT-PCR检测方法，以量化hTERT mRNA的全范围拷贝数。我们在134例已知长期预后的单侧浸润性原发性乳腺癌患者系列中验证了该方法。四分之三的乳腺肿瘤（75.4%；134例中的101例）为hTERT阳性，即含有可检测和可量化的hTERT mRNA。与hTERT阴性患者相比，hTERT阳性患者术后无复发生存期显著缩短（P = 0.017）。在Cox多变量回归分析中，hTERT状态的预后意义仍然存在。当我们将hTERT阳性患者（n = 101）分为三个相等的组（hTERT mRNA含量显示少量、中等或大量增加的肿瘤）时，我们观察到hTERT mRNA高水平与斯卡夫-布鲁姆-理查森组织病理学III级（P = 0.066）、雌激素受体阴性（P = 0.002）和孕激素受体阴性（P = 0.048）状态之间存在统计学关联（或有这种趋势），因此与乳腺肿瘤的更高侵袭性相关。hTERT mRNA高水平也与MYC基因过表达相关（P = 0.007）。这些发现表明，hTERT mRNA的定量评估在人类乳腺癌中可能具有重要的预后意义。此外，我们简单、快速且半自动的检测方法适用于常规hTERT mRNA检测和定量，并且将成为大型随机前瞻性合作组试验以及基于hTERT的治疗项目中的有力工具。
