利用逆转录聚合酶链反应及DNA印迹杂交技术检测乳腺癌腋窝淋巴结微转移

Detection of micrometastases in the axillary lymph nodes in patients with breast cancer by reverse transcriptase polymerase chain reaction and southern hybridization

目的 评价检测原发性乳腺癌腋窝淋巴结微转移的方法。方法 用逆转录聚合酶链反应 (RT PCR)和Southern杂交方法 ,检测腋窝淋巴结中CK 19基因表达。同时与免疫组织化学 (组化 )方法比较其检测敏感性。结果 RT PCR、Southern杂交及免疫组化方法的检测敏感性分别为 1∶5× 10 5、1∶10 6和 1∶5× 10 4 。 5 2例患者的 170枚常规病理检查阴性淋巴结检出微转移 35枚 ,占 2 0 6 %。其中 17例常规病理检查有转移病例的 33枚阴性淋巴结中 ,查出 2 1枚CK 19表达阳性 ;35例淋巴结常规病理检查无转移中有 11例查出微转移 (31 4% )。微转移与其他临床指标无相关性。结论 以CK 19为标志物 ,RT PCR并Southern杂交方法检测原发性乳腺癌淋巴结微转移灵敏、特异 ,可作为临床判断预后的参考指标。

Objective This study was to detect micrometastases in the axillary lymph nodes in primary breast cancer patients and investigate its clinical significance. Methods The cytokeratin 19 gene expression was examined by RT-PCR and Southern hybridization methods. The sensitivity was compared with that of immunohistochemistry(IHC) method.Results The sensitivity was 1∶5×105, 1∶106 and 1∶5×104 by RT-PCR, Southern hybridization and IHC methods,respectively. Thirty-five of 170 routine pathology negative lymph nodes (206%)in 52 primary breast cancer patients contained CK-19 positive signals. Twenty-one out of 33(636%) routine negative lymph nodes were found micrometastases positive in 17 patients who proved to be with lymph node metastases by routine pathologic examination.Eleven out of 35(314%) metastasis negative patients by routine pathology were CK-19 gene positive.Hence the micrometastasis found by our method is an independent parameter.Conclusion RT-PCR method, using CK-19 as a biological marker,was sensitive and specific in detecting micrometastases in primary breast cancer,it could be a new prognostic parameter for breast cancer patients.