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龚丽明, 林世杰, 王忠明, 李鸿钧. 巢式RT-PCR 扩增CK20 、MMP-7 诊断大肠癌淋巴结微转移的初步研究[J]. 肿瘤防治研究, 2005, 32(06): 346-349. DOI: 10.3971/j.issn.1000-8578.837
引用本文: 龚丽明, 林世杰, 王忠明, 李鸿钧. 巢式RT-PCR 扩增CK20 、MMP-7 诊断大肠癌淋巴结微转移的初步研究[J]. 肿瘤防治研究, 2005, 32(06): 346-349. DOI: 10.3971/j.issn.1000-8578.837
GONG Li-ming, LIN Shi-jie, WANG Zhong-ming, LI Hong-jun. Study on Detecting Colorectal Cancer Micrometastases in Lymph Nodes by Nested Reverse Transcriptase-Polymerase Chain Reaction Amplif ication of Cytokeratin 20 and Matrix Metalloprotreinase-7[J]. Cancer Research on Prevention and Treatment, 2005, 32(06): 346-349. DOI: 10.3971/j.issn.1000-8578.837
Citation: GONG Li-ming, LIN Shi-jie, WANG Zhong-ming, LI Hong-jun. Study on Detecting Colorectal Cancer Micrometastases in Lymph Nodes by Nested Reverse Transcriptase-Polymerase Chain Reaction Amplif ication of Cytokeratin 20 and Matrix Metalloprotreinase-7[J]. Cancer Research on Prevention and Treatment, 2005, 32(06): 346-349. DOI: 10.3971/j.issn.1000-8578.837

巢式RT-PCR 扩增CK20 、MMP-7 诊断大肠癌淋巴结微转移的初步研究

Study on Detecting Colorectal Cancer Micrometastases in Lymph Nodes by Nested Reverse Transcriptase-Polymerase Chain Reaction Amplif ication of Cytokeratin 20 and Matrix Metalloprotreinase-7

  • 摘要: 目的 建立巢式逆转录聚合酶链式反应(nested reverse transcriptase-polymerase chain reaction,nested RT-PCR)扩增CK20mRNA、MMP-7mRNA,检测大肠癌淋巴结微转移。方法 据CK20、MMP-7基因序列设计四对引物,建立巢式RT-PCR扩增体系,检测CK20 mRNA、MMP-7 mRNA在30例行根治性手术大肠癌患者187个淋巴结中及10例良性肠道疾病患者25个淋巴结中的表达,并与传统组织学检查相比较。结果 大肠癌患者187个淋巴结中75个CK20 mRNA表达,阳性率40.11%(75/187),97个MMP-7 mRNA表达,阳性率51.87%(97/187),61个CK20 mRNA及MMP-7 mRNA表达,阳性率32.62%,病理组织学检查仅57个淋巴结有肿瘤转移;10例良性肠道疾病25个淋巴结扩增结果阴性。结论 巢式逆转录聚合酶链式反应检测淋巴结CK20 mRNA、MMP-7 mRNA的表达是诊断大肠癌微细胞转移灵敏特异的指标,其结果较常规病理组织学检查敏感,可对精确大肠癌的临床分期、判断患者预后及治疗提供一定理论依据。

     

    Abstract: Objective  To develop a nested reverse t ranscriptase polymerase chain reaction method to amplify Cytokeratin 20mRNA and Matrix Metalloproteinase-7mRNA for detecting colorectal cancer micro-metastases in lymph nodes . Methods  The RT-PCR system was set up by using nest primer. The presence of CK20 mRNA and MMP-7 mRNA was investigated by nested RT-PCR in 100 lymph nodes from 30 colorectal cancer patients who underwent curative operation. Tweenty-five lymph nodes f rom 10 patients with benign intestinal disease were served as negative control. The result s of nested RT-PCR were compared with ones of routine histology. Results  The positive rate of CK20 mRNA and MMP-7 mRNA in 187 lymph nodes from 30 colorectalcancer patient s was 40. 11 %(75/ 187), 51. 87 %(97/ 187) respectively, while only 57 samples were histologically positive. The expression of CK20 mRNA and MMP-7 mRNA in 25 lymph nodes of 10 cases of cont rol group were all negative. Conclusion  Nested reverse transcriptase polymerase chain reaction was a sensitive and specific method to indicate the expression of CK20 mRNA and MMP-7 mRNA in lymph nodes as a helpful marker of micrometastases of colorectal carcinoma cells. It is superior to routine histopathological assay in examing lymph node metastases. The detection of CK20 mRNA and MMP-7 mRNA in lymph nodes may cont ribute to improve the accuracy of clinical staging, determine patient s' prognosis and provide information for adjuvant therapy.

     

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