Quantitative detection of micrometastases in the lymph nodes of gastric cancer patients with real-time RT-PCR: A comparative study with immunohistochemistry
Quantitative detection of micrometastases in the lymph nodes of gastric cancer patients with real-time RT-PCR: A comparative study with immunohistochemistry
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DOI：
10.1002/ijc.11031
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发表时间：
2003-05-20
影响因子：
6.4
通讯作者：
Kaminishi, M
中科院分区：
文献类型：
--
作者：
Kubota, K;Nakanishi, H;Kaminishi, M
组织学检查对发现胃癌淋巴结微转移缺乏敏感性。在本研究中，我们应用实时RT-PCR方法定量检测胃癌淋巴结中的微转移，并与常规组织学和免疫组化的诊断能力进行比较。我们研究了21例接受根治性手术的胃癌患者的392个淋巴结。以癌胚抗原（CEA）和细胞角蛋白-20（CK 20）为标记基因，在LightCycler仪器上进行实时定量RT-PCR。免疫组化与抗体的广泛角蛋白也进行了淋巴结比较的敏感性和特异性。组织学（+）、免疫组化（+）/组织学（-）、免疫组化（-）/组织学（-）和阴性对照结果患者淋巴结中CEA mRNA的中位数（平均）值分别为4，600（16，000）、200（400）、0（9.8）和0（0.6）。由于CEA和CK 20免疫组化显示存在低基因表达的胃癌，因此单纯CEA和CK 20实时RT-PCR存在一些假阴性结果。CEA与CK 20（双重）实时RTPCR的组合部分弥补了这一弱点。因此，所有71个组织学+淋巴结的双重实时RT-PCR以及宽角蛋白免疫组织化学均为阳性。组织学、免疫组化和双重实时荧光定量PCR的阳性率分别为18.0%（71/392）、20.9%（82/392）和25.8%（101/392）。在8例pTINO患者中，2例通过实时RT-PCR观察到阳性淋巴结，但免疫组化未观察到阳性淋巴结。结果表明，双重实时荧光定量RT-PCR是检测胃癌淋巴结微转移最敏感的方法，可用于判断胃癌患者淋巴结微转移的预后。(C)2003 Wiley-Liss，Inc.
Histologic examination lacks the sensitivity to detect micrometastases in gastric cancer lymph nodes. In the present study, we applied a real-time RT-PCR approach to the quantitative detection of micrometastases in gastric cancer lymph nodes and compared diagnostic power with routine histology and immunohistochemistry. We studied 392 lymph nodes from 21 gastric cancer patients who underwent curative surgery. Real-time quantitative RT-PCR was performed on a LightCycler instrument using a hybridization probe for carcinoembryonic antigen (CEA) and cytokeratin-20 (CK20) as marker genes. Immunohistochemistry with antibodies to wide-keratin was also performed in the lymph nodes to compare the sensitivity and specificity. Median (average) values of CEA mRNA in lymph nodes in patients with histology(+), immunohistochemistry(+)/histology(-), immunohistochemistry(-)/histology(-) and negative control results were 4,600 (16,000), 200 (400), 0 (9.8) and 0 (0.6), respectively. There were some false-negative results with simple CEA and CK20 real-time RT-PCR due to the presence of low gene-expressing gastric cancers as revealed by CEA and CK20 immunohistochemistry. CEA in combination with CK20 (duplex) real-time RTPCR partially covered this weakness. Consequently, all 71 histology+ lymph nodes were positive for duplex real-time RT-PCR as well as wide-keratin immunohistochemistry. Positivity rates by histology, wide-keratin immunohistochemistry and duplex real-time RT-PCR were 18.0% (71/392), 20.9% (82/392) and 25.8% (101/392), respectively. In 2 of 8 patients with pTINO, positive lymph nodes were observed by real-time RT-PCR but not by immunohistochemistry. These results indicate that duplex quantitative real-time RT-PCR is the most sensitive method for detecting micrometastases and useful for evaluating the prognostic significance of lymph node micrometastasis in gastric cancer patients. (C) 2003 Wiley-Liss, Inc.