Leone F, Perissinotto E, Viale A, Cavalloni G, Taraglio S, Capaldi A, Piacibello W, Torchio B, Aglietta M
Ordine Mauriziano, Institute for Cancer Research and Treatment, Candiolo, Italy.
Bone Marrow Transplant. 2001 Mar;27(5):517-23. doi: 10.1038/sj.bmt.1702815.
Identification of sensitive techniques for breast cancer cell detection might be relevant for high-dose chemotherapy programs with autologous stem cell transplantation. We investigated the feasibility of Maspin, Mammaglobin and c-ErbB-2 amplification by real-time quantitative polymerase chain reaction (RQ-PCR) for the detection of breast cancer cells in leukaphereses. Expression of the three markers was determined in primary breast cancers and cell lines. Peripheral blood (PB), bone marrow (BM), and leukapheresis samples from patients with malignancies other than breast cancer were used as controls. Sensitivity was evaluated by dilution of primary tumors and cell lines with mononuclear blood cells. We found expression of the three markers in all primary tumors and most cell lines. No blood specimen from control patients had the Maspin transcript, while only one was positive for Mammaglobin. Weak c-ErbB-2 expression was detectable in most PB, all BM and all leukapheresis samples from controls. We observed a low sensitivity of Maspin RQ-PCR and a sensitivity of Mammaglobin RQ-PCR up to one tumor cell in 10(6) mononuclear cells. One out of 18 leukaphereses from breast cancer patients screened for the presence of Mammaglobin mRNA was positive. We conclude that Mammaglobin RQ-PCR might be a useful tool for detection of leukapheresis contamination.
确定用于乳腺癌细胞检测的敏感技术可能与自体干细胞移植的高剂量化疗方案相关。我们研究了通过实时定量聚合酶链反应（RQ-PCR）检测Maspin、乳腺珠蛋白和c-ErbB-2扩增在白细胞分离物中检测乳腺癌细胞的可行性。在原发性乳腺癌和细胞系中测定了这三种标志物的表达。来自非乳腺癌恶性肿瘤患者的外周血（PB）、骨髓（BM）和白细胞分离样本用作对照。通过用单核血细胞稀释原发性肿瘤和细胞系来评估敏感性。我们在所有原发性肿瘤和大多数细胞系中发现了这三种标志物的表达。对照患者的血液标本中没有Maspin转录本，而只有一份乳腺珠蛋白呈阳性。在大多数PB、所有BM和所有对照的白细胞分离样本中可检测到弱的c-ErbB-2表达。我们观察到Maspin RQ-PCR的敏感性较低，而乳腺珠蛋白RQ-PCR在10(6)个单核细胞中检测到一个肿瘤细胞的敏感性较高。在筛查乳腺珠蛋白mRNA的18份乳腺癌患者白细胞分离物中，有一份呈阳性。我们得出结论，乳腺珠蛋白RQ-PCR可能是检测白细胞分离物污染的有用工具。
