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逆转录-聚合酶链式反应检测大肠癌患者淋巴结微转移

Detection of occult metastases in lymph nodes from patients with colorectal carcinoma by reverse transcriptase-polymerase chain reaction

摘要:

目的探讨大肠癌患者淋巴结的微转移及其临床意义.方法采用苏木素-伊红(HE)病理切片染色和细胞角蛋白20(CK20)的RT-PCR 法检测了经病理证实的39例大肠癌患者的260枚淋巴结的肿瘤转移情况.10 枚正常淋巴结为阴性对照,HT29结肠癌细胞和5例大肠癌标本为阳性对照.结果 10枚正常淋巴结CK20表达阴性,HT29细胞和大肠癌标本CK20表达阳性.HE 染色查见转移的1 6例患者的29枚淋巴结CK20 mRNA全部阳性,而在组织学未见转移的淋巴结中,有5例患者的28枚呈CK20阳性表达,即存在癌转移,显示两种方法的检测结果差异显著(11.1%和21.9 %,P<0.01).按照HE染色结果,Dukes' A、B、 C和D期的病例数分别为3、20、12和4 例,在20例Dukes' B期患者中,5例有CK20表达阳性的淋巴结.结论 CK20的RT-PCR检测是发现肿瘤淋巴结微转移的敏感、特异和简便的方法.大肠癌患者淋巴结CK20 mRNA 的检测可用于大肠癌的准确分期和围手术期辅助治疗方案的选择,但需进一步研究加以证实.

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Objective To detect occult metastases in lymph nodes from patients with colorectal carcinoma and its clinical significance.Methods The metastases in 260 lymph nodes from 39 histologically verified colorectal cancer patients were studied by both hematoxylin-eosin (HE) staining and cytokeratin-20 (CK20) specific RT-PCR. Ten normal lymph nodes were served as negative controls, and HT29 colon cancer cell line and 5 colorectal cancer specimens as positive controls. Results Ten normal lymph nodes were CK20-negative, HT29 cells and 5 tumor specimens were all CK20-positive. All 29 lymph nodes from 16 patients which confirmed metastases by HE staining exhibited CK20 positive expression; an additional 28 lymph nodes from 5 patients with no histologically detectable metastases expressed CK20 mRNA, i.e. presence of metastases. The difference of the positivity was significant (11.1% vs 21.9%, P<0.01). According to the HE staining, the cases of Dukes' A, B, C and D were 3, 20, 12 and 4, respectively. In the 20 patients of Dukes' B stage, 5 of them had CK20-positive lymph nodes. Conclusion CK20-specific RT-PCR is a highly sensitive, specific and simple method for detecting occult metastases in lymph nodes. The detection of CK20 mRNA expression in lymph nodes is recommended to precisely determine tumor stage and postoperative adjuvant therapy for patients with colorectal cancer, and further studies should be done in future to confirm the findings.

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