Detection of nonmelanoma skin cancer micrometastases in lymph nodes by using reverse transcriptase-polymerase chain reaction for keratin 19 mRNA
Detection of nonmelanoma skin cancer micrometastases in lymph nodes by using reverse transcriptase-polymerase chain reaction for keratin 19 mRNA
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DOI：
10.1111/j.1365-2133.2003.05602.x
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发表时间：
2003-11-01
影响因子：
10.3
通讯作者：
Kitajima, Y
中科院分区：
文献类型：
--
作者：
Kamiya, M;Ichiki, Y;Kitajima, Y
背景应用逆转录-聚合酶链反应（RT-PCR）扩增角蛋白19（K19）mRNA是检测皮肤癌淋巴结微转移的一种新的敏感方法。目的比较RT-PCR与组织学检查对皮肤癌微转移的检出率。方法对13例皮肤鳞状细胞癌患者的26枚淋巴结（26枚淋巴结）进行RT-PCR检测，并与病理组织学检查结果进行比较。SCC）、小汗腺汗孔癌和佩吉特病进行组织学检查（苏木精-伊红切片）和RT-PCR。用K19引物对提取的RNA进行RT-PCR。RT-PCR产物经溴化乙锭染色后，与K19特异性探针进行非放射性杂交。在16个组织学阴性的淋巴结中，通过RT-PCR发现1个（6%）表达K19 mRNA，表明存在组织学检查无法检测到的微转移。使用从SCC细胞提取的RNA与从正常淋巴结细胞提取的RNA混合的系列稀释研究显示，K19 RT-PCR的检测灵敏度为1 μ g淋巴结RNA中10（-5）μ g癌细胞RNA。巢式RT-PCR检测敏感性为1/10（6）淋巴细胞。结论K19 RT-PCR检测皮肤癌淋巴结微转移是有效的。
Background A new sensitive method for the detection of skin cancer micrometastases in lymph nodes is based on amplification of keratin 19 (K19) mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR).Objectives To compare results of RT-PCR with those of histological examination in terms of the detection rate of skin cancer micrometastases.Methods Twenty-six lymph nodes obtained from 13 patients with squamous cell carcinoma (SCC), eccrine porocarcinoma and Paget's disease were investigated by histological examination (haematoxylin and eosin sections) and RT-PCR. RT-PCR was performed on extracted RNA by using K19 primer pairs. RT-PCR products were visualized by ethidium bromide staining and confirmed by non-radioactive hybridization with K19-specific probes.Results All of 10 histologically positive lymph nodes yielded the expected 460-bp band. Of the 16 histologically negative lymph nodes, one (6%) was found by RT-PCR to express K19 mRNA, indicating the presence of micrometastases which could not be detected by histological examination. A serial dilution study using RNA extracted from SCC cells mixed with RNA extracted from normal lymph node cells showed a detection sensitivity of K19 RT-PCR of 10(-5) mug cancer cell RNA in 1 mug lymph node RNA. Nested RT-PCR showed a detection sensitivity of one tumour cell in 10(6) lymphocytes.Conclusions These results demonstrate the usefulness of K19 RT-PCR for the detection of skin cancer micrometastases in lymph nodes.