CK19和hMAM及SBEM检测乳腺癌微转移价值分析

目的：探讨生物学标志乳腺上皮小黏蛋白（smallbreastepithelialmucin，SBEM）、乳腺珠蛋白（humanmamma—globin，hMAM）和细胞角蛋白19（cytokeratin19，CK19）检测乳腺癌外周血及淋巴结微转移的临床应用价值。方法：收集2012-04—13—2012-10—10广西医科大学附属肿瘤医院乳腺外科首诊的患者外周血及组织标本。采用巢式PCR技术对47例乳腺癌患者和45例乳腺良性肿瘤患者外周血有核细胞中SBEM、hMAM和CK19的mRNA进行检测；采用半定量PCR技术对10例乳腺癌组织及对应的癌旁组织，以及10例乳腺良性肿瘤患者组织mRMA进行检测；采用免疫组化技术对其中27例乳腺癌患者的109个淋巴结进行检测。结果：乳腺癌和乳腺良性肿瘤患者的外周血SBEM表达率分别为82．98％（39／47）和62．22％（28／45），χ2=5．005，P=0．021；乳腺癌和乳腺良性肿瘤患者的外周血hMAM表达率分别为65．96％（31／47）和42．22％（19／45），差异有统计学意义，χ2=5．220，P=0．022；乳腺癌和乳腺良性肿瘤患者的外周血CK19表达率分别为89．36％（42／47）和80．00％（36／45），差异无统计学意义，χ2=1．562，P=0．211。免疫组化CK19在52个转移性淋巴结中强阳性表达，在57个常规病理检查未发现的转移性淋巴结中检出5个淋巴结存在转移灶；hMAM及SBEM在转移性淋巴结中表达比CK19弱，而且阳性细胞染色不均一。结论：SBEM、hMAM和CK19作为乳腺癌外周血微转移生物学标志具有较高的灵敏性，但缺乏特异性，联合检测可以提高特异性，对发现早期乳腺癌微转移有一定临床意义；CK19可用于检测乳腺癌腋窝淋巴结的微小转移灶。

Micrometastasis detection in breast cancer of patients by biological markers CK19 ,hMAM and SBEM

OBJECTIVE： To disscus the clinical application of biological markers small breast epithelial mucin,human mammaglobin,and cytokeratin19 by detecting circulating tumor cells and lymph node micrometastasis in breast cancer. METHODS：The peripheral blood and tissue samples were collected in diagnosed patients of breast cancer from April 13, 2012 to October 10, 2012, Department of Breast Surgery, Tumor Hospital Affiliated to Guangxi Medical University. SBEM,hMAM,and CK19 mRNA were dected by nested P CR in peripheral blood of 47 eases of breast cancer and 45 eases of benign breast tumor. Semi-quantitative PCR method was used to detected the mRNA express of 10 cases of breast canc- er tissue, 10 cases of paraeancerous tissue and 10 cases of benign breast tumor tissue. The metastasis of 109 lymph nodes in 27 cases of breast cancer patients were detected using immunohistochemistry technique. RESULTS：SBEM expression in peripheral blood of breast cancer and benign breast tumor patients were 82.98 % （ 39/47） and 62.22 % （28/45）, respectively （χ2=5. 005,P--0. 021）;hMAM expression in peripheral blood of breast cancer and benign breast tumor patients were 65.96%（31/47） and 42.22% （ 19/45）, respectively （χ2= = 5. 220, P= 0. 022）, the difference was statiscally significant. CK19 expression in peripheral blood of breast cancer and benign breast tumor patients were 89.36% （42/47） and 80.00 （36/45）, respectively （χ2== 1. 562 ,P=0.211 ）, the difference was no statisca significace. Fifty-two lymph nodes metastasis by HE staining,there had 5 lymph nodes micometastasis were found by CK19 in 57 negative lympb nodes diagnosed by routine pathology, hMAM and SBEM expression in metastatic lymph nodes was weaker than that of CK19 and the positive cells were not uniform. CONCLUSIONS= SBEM, hMAM,and CK19 as biomarkers for breast cancer micrommastases in pe ripheral blood have high sensitivity and low specificity. Joint detection could improve the specificity and it has some clinical significance for the early detection of breast cancer micrometastases. CK19 may be used to detect axillary lymph node mb crometastases.