Oral cancer diagnosis via a ferrocenylnaphthalene diimide-based electrochemical telomerase assay.
Oral cancer diagnosis via a ferrocenylnaphthalene diimide-based electrochemical telomerase assay.
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DOI：
10.1373/clinchem.2012.191569
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发表时间：
2013
影响因子：
9.3
通讯作者：
Kumiko Mori;Shinobu Sato;M. Kodama;M. Habu;O. Takahashi;T. Nishihara;K. Tominaga;S. Takenaka
中科院分区：
文献类型：
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作者：
Kumiko Mori;Shinobu Sato;M. Kodama;M. Habu;O. Takahashi;T. Nishihara;K. Tominaga;S. Takenaka
端粒酶被认为是一个很好的癌症标志物，因为与正常细胞相比，它在大多数肿瘤细胞中不受调节。我们通过电化学技术评估了口腔癌患者肿瘤组织和周围细胞裂解液中端粒酶的活性，称为电化学端粒酶测定（ECTA）。方法以二茂铁萘二亚胺（FND）为探针。电极上固定的电化学端粒酶底物（ETS）引物被端粒酶拉长，与产物结合的FND产生电流。数据分别用拉伸前后电流的变化进行了标准化。结果：在大多数癌症患者的活检样本中，电流的变化增加了30%以上，而在大多数健康个体中，电流的变化增加了20%或更低。在这种差异的基础上，个体临床样本被判断端粒酶阳性、模糊或阴性。端粒酶重复扩增法在癌组织和脱落细胞中的阳性率分别为85%和90%，而端粒酶重复扩增法在癌组织和脱落细胞中的阳性率分别为50%和10%。此外，ECTA在小于2 cm的早期肿瘤中阳性率为100%，健康人活检细胞和脱落细胞的阳性率分别为95%和82%。56份未知的EOCs样本在84%的病例中被正确判断为癌症或正常。结论ECTA对癌细胞和正常细胞均有较高的检测准确率，尤其对EOCs的检测准确率较高，表明该微创检测方法适用于口腔癌的诊断。
BACKGROUND Telomerase is regarded as a good marker for cancer because it is unregulated in most tumor cells compared with normal cells. We evaluated telomerase activity in the lysate of tumor tissue and surrounding cells of oral cancer patients by an electrochemical technique, dubbed the electrochemical telomerase assay (ECTA). METHODS The assay used ferrocenylnaphthalene diimide (FND) as the probe. Electrochemical telomerase substrate (ETS) primer immobilized on the electrode was elongated by telomerase and FND bound to the product to give rise to a current. The data were standardized with the change in current before and after the elongation, respectively. RESULTS The change in current increased more than 30% in biopsy samples from most cancer patients, whereas the increase was 20% or lower in most healthy individuals. On the basis of this difference individual clinical samples were judged telomerase positive, ambiguous, or negative. The positive rates in the cancerous tissues and exfoliated cells (EOCs) of the patients were 85% and 90%, respectively, whereas the corresponding values were 50% and 10% by the telomerase repeat amplification protocol. Furthermore, the positive rate for the ECTA was 100% in early tumors smaller than 2 cm, and 95% and 82% of biopsy and exfoliated cells of healthy individuals were correctly judged as negative. Fifty-six unknown samples with EOCs tested were correctly judged to be cancerous or normal in 84% of cases. CONCLUSIONS The ECTA yielded high hit rates for cancerous and normal cells, especially in EOCs, results indicating that this minimally invasive test is suitable for oral cancer diagnosis.