Taniyama Kiyomi, Motoshita Junichi, Sakane Junichi, Makita Kyoko, Akai Yasumasa, Daito Motonari, Otomo Yasuhiro, Ono Hiroshi, Mizunoe Tomoya, Takeuchi Yukiyasu, Tominaga Harumi, Koseki Masato
Institute for Clinical Research, Department of Pathology, National Hospital Organization Kure Medical Center and Chugoku Cancer Center, Kure, Japan.
Pathobiology. 2006;73(4):183-91. doi: 10.1159/000096019.
The aim was to develop a more efficient molecular detection system than histological examination (HE) for lymph node (LN) metastasis.
Cytokeratin (CK) 19 mRNA copy numbers of 5 colon carcinoma cell lines (Lovo, DLD1, WiDr, Colo201 and Colo320) were calculated and compared by one-step nucleic acid amplification (OSNA) and conventional real-time reverse-transcription polymerase chain reaction (RT-PCR). Then, 91 LN submitted for HE from 6 patients with advanced colorectal adenocarcinoma and 64 LN submitted for frozen diagnosis from 47 patients with different malignancies were examined by OSNA and HE.
CK19 mRNA copy numbers of all but Colo320 cells detected by OSNA were within double of those detected by RT-PCR. The least cell count of Lovo cells detected at one reaction (2 microl) by OSNA was calculated as 0.8 cells. Carcinoma metastasis showing either HE+ or OSNA+ was detected in 7.9% of the LN from advanced colorectal adenocarcinomas and in 30.0% of the LN for frozen diagnosis from different malignancies; HE-/OSNA+ metastasis was detected in 4.8 and 4.0%, respectively. OSNA analysis of 1 LN could be completed within 40 min.
A combined analysis of LN by HE and OSNA could increase the sensitivity for detecting micrometastasis during surgery.
开发一种比组织学检查（HE）更高效的用于检测淋巴结（LN）转移的分子检测系统。
通过一步核酸扩增（OSNA）和传统实时逆转录聚合酶链反应（RT-PCR）计算并比较5种结肠癌细胞系（Lovo、DLD1、WiDr、Colo201和Colo320）的细胞角蛋白（CK）19 mRNA拷贝数。然后，对6例晚期结直肠癌患者送检HE的91个淋巴结以及47例不同恶性肿瘤患者送检冷冻诊断的64个淋巴结进行OSNA和HE检查。
除Colo320细胞外，OSNA检测的所有细胞系的CK19 mRNA拷贝数均在RT-PCR检测值的两倍以内。OSNA在一次反应（2微升）中检测到的Lovo细胞的最低细胞数经计算为0.8个细胞。在晚期结直肠癌患者的淋巴结中，HE或OSNA检测到癌转移的比例为7.9%，在不同恶性肿瘤冷冻诊断的淋巴结中这一比例为30.0%；HE阴性/OSNA阳性转移分别在4.8%和4.0%的淋巴结中被检测到。对1个淋巴结进行OSNA分析可在40分钟内完成。
HE和OSNA联合分析淋巴结可提高手术中检测微转移的敏感性。
