乳腺癌前哨淋巴结和骨髓和外周血微小转移的联合检测

Combined detection on micrometastases in sentinel lymph node, peripheral blood and bone marrow from breast cancer patients

目的:探讨采用HE染色、免疫组织化学(IHC)和RTPCR等不同方法同时检测乳腺癌前哨淋巴结(SLN)、骨髓和外周血微小转移的灵敏度及其临床意义。方法:全身麻醉后先行骨髓穿刺和外周血采集,选用1%异硫蓝行前哨淋巴结活检(SLNB)后,采用HE染色、IHC和RTPCR等不同方法同时检测乳腺癌SLN、骨髓和外周血的微小转移。结果:腋窝淋巴结常规HE染色病理检查阴性患者38例,4例RTPCR技术和IHC方法检测SLN微小转移同时阳性,另外RTPCR技术还有6例KT19mRNA也表达,两种方法间显著相关,P=0003,但灵敏度上差异无统计学意义,P=0076;骨髓KT19mRNA阳性表达11例,明显高于外周血阳性表达3例,P=0018;RTPCR技术11例骨髓阳性表达,其中6例IHC检测也为阳性,二者间显著相关,P=0000,但灵敏度上差异无统计学意义,P=0169;38例骨髓和SLN中仅2例同时表达KT19mRNA,其间无显著相关,P=0690。结论:即使术前常规检查未发现腋窝淋巴结及远处转移,骨髓、外周血和淋巴结也可检出微小转移灶。由于骨髓和腋窝淋巴结不是同步出现,因此临床上需要检测多种组织、多个指标,才能更精确地对微小转移进行评价。

OBJECTIVE:To study the value of hematoxylin and eosin (HE) staining, immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR) in the detection of micrometastases in sentinel lymph node (SLN), peripheral blood and bone marrow samples from breast cancer patients and the clinical significance of micrometastases. METHODS:After general anesthetized, all patients were performed sentinel lymph node biopsy (SLNB) by 1% isosulfan blue. Micrometastases were detected by HE staining, IHC and RT-PCR in SLN, peripheral blood and bone marrow samples from 38 breast cancer patients.RESULTS:Of the 38 paired bone marrow and peripheral blood samples, 11 bone marrow samples (28.95%) were positive by RT-PCR, whereas only 3 (7.89%) peripheral blood samples were positive (χ2=5.604, P=0.018). Comparing different methods in detection of bone marrow micrometastases, 6 (15.79%) were IHC positive and 11 (28.95%) were RT-PCR positive. But there was no significant difference statistically (χ2=1.894, P=0.169) between the two methods. Of the 11 bone marrow micrometastases cases detected by RT-PCR, 6 were IHC positive. A correlation was showed statistically between the method of RT-PCR and IHC ( P=0.000, r=0.678). In SLN samples, 10 (26.32%) were RT-PCR positive, while 4 (10.53%) were IHC positive. But there is no statistical difference between RT-PCR and IHC (χ2=3.152, P=0.076). 4 SLN samples were both RT-PCR positive and IHC positive, else 6 SLN samples were only RT-PCR positive. A correlation was showed between RT-PCR and IHC methods (P=0.003, r=0.574). 2 of 38 paired bone marrow and SLN samples were detected both positive by RT-PCR, which indicated that bone marrow micrometastases did not always simultaneously occur with SLN micrometastases(P=0.690). CONCLUSIONS:Micrometastases can be detected in bone marrow, peripheral blood and lymph nodes samples from patients who reveal no distant metastases by preoperatively regular examination. Because micrometastases are not always simultaneously detected in different tissues, it is important to detect some tissues and adopt some factors so that micrometastases status can be evaluated correctly and effectively.