毛细管电泳检测nm23-H_1基因在乳腺癌组织中的表达

Expression of nm23-H_1 in Breast Cancer Detected by Capillary Electrophoresis

目的:检测nm23-H1基因在乳腺癌中的表达与突变,寻找它们与淋巴结转移的关系。方法:用半定量逆转录-聚合酶链反应(RT-PCR)检测nm23-H1mRNA表达,毛细管聚丙烯酰胺凝胶电泳(CPAGE)行单链构象多态性分析(SSCP)检测DNA突变。结果:应用CPAGE可在30min内分离出nm23-H1基因RT-PCR产物的单链及双链峰。nm23-H1表达、突变在腋淋巴结有、无转移组均有差异;mRNA在有远处转移组呈现低表达;DNA突变率在肿瘤≤5cm和>5cm组间有差异。结论:nm23-H1表达和突变可以做为评估乳腺癌转移危险的指标;CPAGE技术用于肿瘤组织的PCR-SSCP分析具有快速灵敏、准确、重复性好的特点。

Objective: To examine the expression and mutation of nm23-H1 gene in breast cancer,and to detect the relationship between them and lymph node metastasis. Methods: The expression of nm23-H1 mRNA was detected by Reverse Transcription-Polymerase Chain Reaction (RT- PCR),and DNA mutation was detected by Capillary Polyacrylamide Gel Electrophresis (CPAGE) and Single Strand Conformation Polymorphism (SSCP) analysis. Results: nm23-H1 gene RT-PCR production could be separated into dsDNA peak and ssDNA peaks within 30 minutes by CPAGE.nm23-H1 expression and mutation were significantly different between patients with and without lymph node metastasis, and the expression of mRNA was low in distant metastasis group. DNA mutation showed a significant difference between tumor smaller than 5 cm in size and greater than 5 cm in size. Conclusion: The expression and mutation of nm23-H1 gene in breast cancer can serve as indexes for scanning the risk of metastasis. CPAGE technique is sensitive, accurate and well reproducible in tumor tissue PCR-SSCP analysis.