乳腺癌骨髓微小转移灶的检测与意义

骨是乳腺癌最常见的转移部位之一，大约30％左右的可手术乳腺癌存在骨髓微小转移灶。主要阐述了乳腺癌骨髓微小转移灶的检测方法及其预后意义，同时对乳腺癌微小转移灶的生物学特性和对辅助治疗的反应加以综述。     

乳腺癌骨髓微小转移灶检测的临床意义

本文综述了乳腺癌骨髓微小转移灶早期检测的研究状况，着重介绍了利用免疫组化技术及ＰＣＲ技术检测早期乳腺癌骨髓中的微小转移灶，并结合近年来押外文献，探讨其与乳腺癌预后的关系。     

抗转移基因nm23在人乳腺癌中的表达

肿瘤转移是癌症忠者的主要死亡原因,早期乳腺癌患者进行骨髓检查约28—35%有微小转移灶,40～50%发生转移。精确估计原发瘤的转移能力和未检出的转移灶对乳腺癌患者的处理是十分重要的。肿瘤转移涉及许多过程,包括有特殊特性的肿瘤细胞在宿主体内生长并建立转移灶。由     

54例乳腺癌患者骨髓象分析

５４例乳腺癌患者骨髓象分析张奇威，罗健松，崔景华，邵植辉广州中山医科大学肿瘤医院内科实验室（５１００６０）乳腺癌患者骨髓象能否确定乳癌有无微小转移灶，目前罕见报道。现将１９８２年１月至１９９０年３月收集乳腺癌５４例报道如下，试论骨髓检查在乳癌诊断中的...     

乳腺癌前哨淋巴结微小转移灶的预后意义及临床处理

随着乳腺癌前哨淋巴结活组织检查技术的广泛开展与病理诊断水平的不断提高,在临床工作中,乳腺癌前哨淋巴结微小转移灶的检出率明显增加。对于乳腺癌前哨淋巴结微小转移灶患者预后影响,目前国内外尚未达成一致意见,部分研究结果显示其预后意义与腋窝淋巴结清扫术后查见微小转移灶不尽相同。前哨淋巴结微转移患者的最佳临床决策目前亦无定论,当前多数研究倾向于对这类患者可采用全身辅助治疗联合术后局部放射治疗取代腋窝淋巴结清扫术,但该结论仍需进一步临床研究证实。笔者复习近年来国内外相关文献,对乳腺癌前哨淋巴结微小转移灶的预后意义及临床处理方法作一综述。     

腋窝淋巴结微小转移与肿瘤微血管计数对乳腺癌患者预后的影响

目的:通过对腋窝淋巴结阴性乳腺癌患者的淋巴结进一步分析,研究淋巴结微小转移灶及肿瘤微血管计数(MVD)对预后的影响.方法:选取1993年564例乳腺癌中腋窝淋巴结阴性患者48例(24例死亡,24例生存),分别用HE、EMA和CK19对原淋巴结病理切片进行复染,确定微小转移灶.将肿瘤病理切片用FVIII因子染色,确定微血管数目.结果:48例共882枚淋巴结中发现微小转移灶为9.0%(79/882),死亡组与对照组之间无显著差异(P＞0.05);微血管计数两组之间差异有显著性意义(P＜0.001),经统计学分析微血管计数对患者预后的判别能力大于淋巴结微小转移灶.结论:本研究中淋巴结微小转移灶未显示出对生存的明显影响,而肿瘤的微血管计数与腋窝淋巴结阴性乳腺癌患者的生存时间呈负相关.     

乳腺癌前哨淋巴结和骨髓和外周血微小转移的联合检测

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。     

乳腺癌骨髓和外周血微小转移灶的研究进展

乳腺癌是我国女性常见的恶性肿瘤,且发病率呈逐年增高趋势。尽管乳腺癌在早期诊断和综合治疗方面都有了很大的进步,但肿瘤远处转移仍严重影响患者的预后[1]。本文对目前乳腺癌微小转移灶的研究状况做一综述,并结合近年来的有关国外文献,探讨微小转移灶的临床意义。自从Ash-worth首次证实在血液中发现癌细胞以来,人们将它称为肿瘤细胞微小转移病灶(micrometastases)、微小残存病灶(min-imal residual disease,MRD)、循环肿瘤细胞(circulating tumorcells,CTCs)、潜伏肿瘤细胞(occult tumor cells,OTCs)等。     

乳腺癌前哨淋巴结和骨髓和外周血微小转移的联合检测

目的：探讨采用HE染色、免疫组织化学(IHC)和RT—PCR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RT—PCR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RT-PCR技术和IHC方法检测SLN微小转移同时阳性．另外RT-PCR技术还有6例KT19 mRNA也表达，两种方法间显著相关．P=0．003，但灵敏度上差异无统计学意义．P=0．076；骨髓KT19 mRNA阳性表达11例．明显高于外周血阳性表达3例，P=0．018；RT-PCR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=0．000．但灵敏度上差异无统计学意义．P=0．169；38例骨髓和SLN中仅2例同时表达KT19 mRNA，其间无显著相关，P=0．690。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。     

免疫组织化学和RT-PCR法检测乳腺癌骨髓和前哨淋巴结微小转移的临床研究

目的探讨免疫组织化学(IHC)和RT-PCR法检测乳腺癌骨髓和前哨淋巴结(SLN)微小转移的灵敏度及临床意义。方法留取乳腺癌改良根治术腋窝淋巴结HE染色证实阴性的病人的胸骨骨髓血和SLN,分别采用IHC和RT-PCR方法检测其微小转移情况。结果62例中,骨髓样本RT-PCR检测15例阳性表达,其中9例IHC检测也为阳性,二者结果有较好一致性(kappa=0.6945),检出率有统计学差异(P=0.0412);SLN样本RT-PCR法有13例KT19mRNA表达,其中7例IHC检出KT19阳性细胞,二者结果一致性较好(kappa=0.6483),检出率有统计学差异(P=0.0412);骨髓和SLN同时表达KT19mRNA仅3例,无显著相关(P=0.796);原发肿瘤大小和骨髓KT19mRNA表达率有关联(P=0.003)。结论常规检查未发现远处和腋窝淋巴结转移,骨髓和SLN可检出微小转移,RT-PCR较IHC更灵敏,肿瘤大小与骨髓微小转移有关联。由于骨髓和腋窝淋巴结微小转移不一定同步出现,选用灵敏方法对不同组织同时进行检测可能更具临床价值。     

以原因不明发热为主要表现的乳腺癌骨髓转移一例及诊断思维分析

目的探讨乳腺癌骨髓转移的临床特点及诊断思维。方法报告1例乳腺癌骨髓转移病例,收集临床资料并结合临床判断和处理等方面存在的缺陷复习相关文献。 结果患者以不明原因发热为首发症状,通过空芯针穿刺明确为乳腺恶性肿瘤。但患者持续高热,多种抗生素治疗无效,用药期间出现血小板及血红蛋白进行性降低及全身骨痛。后经骨扫描、骨髓穿刺活检诊断为乳腺癌骨髓转移,予单周紫杉醇联合卡铂化疗1疗程后体温正常。结论乳腺癌骨髓转移早期易被忽视,当有骨痛并突发原因不明的贫血和血小板减少时,应考虑骨髓转移,骨髓穿刺是简单而有效的确诊手段。     

Increased serum interleukin-8 in patients with early and metastatic breast cancer correlates with early dissemination and survival.

PURPOSE: The prognostic significance of serum interleukin (IL)-8 was evaluated in patients with metastatic breast cancer. The predictive value of serum IL-8 for the presence of occult metastatic tumor cells in bone marrow aspirates was evaluated in patients with operable and metastatic breast cancer. EXPERIMENTAL DESIGN: Serum IL-8 was measured in healthy controls, patients with operable breast cancer, and patients with untreated, progressive metastatic breast cancer. In 69 patients with either operable or advanced breast cancer, occult cytokeratin-positive cells were counted in bone marrow aspirates. RESULTS: Serum IL-8 levels are increased in 67% (52 of 77) of patients with advanced breast cancer. Overall, these levels are significantly higher in patients with breast cancer compared with healthy volunteers (P < 0.001). The IL-8 levels increase significantly in patients with more advanced disease. An elevated serum IL-8 is related to an accelerated clinical course, a higher tumor load, and the presence of liver or lymph node involvement. A multivariate analysis indicates that serum IL-8 is an independent significant factor for postrelapse survival. There was a significant difference between serum IL-8 levels in patients with or without occult cytokeratin-positive bone marrow cells (P < 0.04). Serum IL-8 levels also showed an association with the number of these cells (P < 0.01). CONCLUSIONS: Serum IL-8 is increased in patients with breast cancer and has an independent prognostic significance for postrelapse survival. The observations on the relationship between occult cytokeratin-positive bone marrow cells corroborate the concept of IL-8 acting as a contributor to the process of tumor cell dissemination. Similarly, the relationship between serum IL-8 and nodal stage at presentation deserves further study. These results further expand the concept that inflammation and inflammatory cytokines are critical components of tumor progression.     

Immunohistochemical staining of bone marrow biopsies for detection of occult metastasis in breast cancer

Summary Immunohistochemical (IHC) techniques should allow for a greater detection of bone marrow micrometastasis in patients with breast carcinoma. We studied a series of bone marrow (BM) biopsies negative by conventional histologic techniques from 93 patients with breast carcinoma. Prior to this study, twelve BM biopsies, positive by conventional histology, were stained with a panel of monoclonal antibodies (MoAb), directed either against cytokeratin (KL1, AE1-AE3, CAM5-2) or epithelial membrane antigen (EMA, HMFG2). KL1 appeared to be the most sensitive of the markers used in the detection of metastases and is available commercially. It therefore was the only MoAb used with the series of 93 BM biopsies negative by conventional examination. Within this series, among 45 patients clinically suspected of having bone marrow metastasis but with BM biopsies negative by conventional staining, one case showing myelofibrosis stained positive with KL1 demonstrating isolated tumor cells. For the 48 patients without suspicion of bone marrow metastasis at initial diagnosis for breast carcinoma, KL1 revealed no marrow metastasis.Single bone marrow biopsy techniques whether stained by conventional or IHC methods do not appear to be useful tests to detect occult bone marrow metastasis, especially at initial diagnosis of clinically M_o breast carcinoma patients.     

Angiogenesis in the bone marrow of patients with breast cancer.

PURPOSE: Pathologic angiogenesis has been correlated with tumor growth, dissemination, metastasis, and prognosis in solid tumors including breast cancer. Angiogenesis has also been implicated in the pathophysiology of, and shown to be a therapeutic target in tumors arising in the bone marrow. The status of angiogenesis in the bone marrow of breast cancer patients is unknown. The aim of this study was to estimate the extent of bone marrow angiogenesis in this subset of patients. EXPERIMENTAL DESIGN: We studied 42 women with breast cancer in whom a bone marrow biopsy was done. Bone marrow samples were sorted according to their infiltration status by breast cancer cells. In all bone marrow sections, blood vessels were highlighted by staining endothelial cells with an antibody directed against the CD34-related antigen. A hematopathologist blind to the status of infiltration of breast cancer did the bone marrow vessel count. RESULTS: Nineteen patients (45%) had bone marrow metastasis. The bone marrow microvessel density was significantly higher in patients with bone marrow metastases compared with patients without bone marrow metastases (P < 0.0005). Median bone marrow microvessel density was 2 for the negative bone marrow group, and 15 for the positive bone marrow group. An increased microvessel density was correlated with presence of disease at last follow-up. CONCLUSIONS: This is the first study showing that bone marrow microvessel density is significantly higher in breast cancer patients with bone marrow metastases, when compared with breast cancer patients without evidence of bone marrow metastatic disease. Further research is needed to shed light into the prognostic and therapeutic relevance of this finding.     

Comparison of monoclonal antibodies for the detection of occult breast carcinoma metastases in bone marrow

Summary Twenty percent (n = 6) of Stage III or IV breast cancer patients (n = 30) had bone marrow metastases detected in bilateral bone marrow biopsy/aspiration preparations using standard histologic preparations. Each metastasis was also detected by four separate monoclonal antibodies (MAbs) which recognize breast carcinoma associated antigens (DF3, anti-EMA, HMFG-2, and CAM5.2). These MAbs were then utilized to stain other bone marrow preparations (n = 81) to determine their utility for the detection of micrometastatic breast carcinoma. MAbs HMFG-2, anti-EMA, and DF3 were each strongly reactive with bone marrows containing histologically-evident metastatic breast carcinoma (18/18). These anti-epithelial membrane antigen MAbs, however, were also reactive with rare plasma cells and immature cells (as well as cell clusters) in some of the control bone marrow samples tested, including those from normal patients and patients with hematologic disorders. They also reacted with some of the preparations from patients with leukemia and lymphoma, and with uninvolved marrows from patients with non-epithelial malignancies. The anti-keratin MAb CAM5.2, in contrast, reacted with 83% (15/18) breast cancer metastases and failed to stain any cells in the various categories of control marrow preparations. These data suggested that MAb CAM5.2 might be utilized to immunohistochemically differentiate micrometastatic breast carcinoma from immature myeloid or erythroid elements.Each MAb was then reacted with histologically uninvolved marrow preparations from the remaining 24 of 30 breast cancer patients in an attempt to identify occult breast carcinoma metastases. While MAbs HMFG-2, DF3, and anti-EMA demonstrated reactive cells in some of these marrows, this reactivity was similar to that seen with control preparations. MAb CAM5.2, in contrast, was negative with all specimens. These data suggest that MAb CAM5.2 may be a useful immunologic probe for the detection and confirmation of metastatic breast carcinoma in bone marrow, while more caution must be employed in the interpretation of results obtained using MAbs anti-EMA, DF3, and HMFG-2.     

检测乳腺癌患者骨髓中CK19的表达及其临床意义

背景与目的:乳腺癌早期即可发生播散和转移,远处转移尤其骨转移是影响患者预后的主要因素之一,而骨髓微转移用常规方法又难以检测出来。本研究旨在探讨检测乳腺癌患者骨髓中CK19mRNA的表达及其临床意义。方法:应用RT鄄PCR技术,同时对照检测65例乳腺癌患者、15例乳腺良性病变患者和8例健康人骨髓中的CK19mRNA表达,并分析CK19mRNA表达与临床病理因素的关系。结果:65例乳腺癌患者中,22例CK19mRNA阳性,阳性率为33.8%;乳腺良性病变患者和健康人骨髓中未检测到CK19mRNA表达。CK19mRNA表达与乳腺癌肿瘤大小和分期有关(P<0.05),与年龄和淋巴结转移状况无显著性相关(P>0.05);与外周血中CEA异常增高呈正相关(r=0.372,P=0.002)。结论:CK19mRNA可作为检测乳腺癌患者骨髓微转移的分子指标之一,可为乳腺癌患者的治疗和预后判断提供帮助。     

隐匿性乳腺癌的诊断和治疗

目的 探讨隐匿性乳腺癌的诊断、治疗及预后.方法 收集23例隐匿性乳腺癌患者的临床和随访资料,并进行分析.结果 23例患者均为女性,平均57.7岁.术前行影像学检查22例,其中行乳腺超声检查17例,8例发现可疑结节;行乳腺钼靶摄片9例,阳性3例;行乳腺MRI检查2例,1例发现异常钙化.20例行同侧乳腺癌改良根治术,16例进行化疗,4例放疗.随访期间,2例患者发生肺转移,其中1例多处转移.结论 术前排除其他部位原发癌的可能后,表现为腋窝淋巴结转移癌的患者即可诊断隐匿性乳腺癌.对隐匿性乳腺癌,乳腺必须进行治疗,可行乳腺癌改良根治术或腋窝淋巴结清扫+全乳放疗.     

A novel HSV-1 virus, JS1/34.5-/47-, purges contaminating breast cancer cells from bone marrow.

PURPOSE: Oncolytic herpes simplex virus type 1 (HSV-1) vectors show considerable promise as agents for cancer therapy. We have developed a novel recombinant HSV-1 virus (JS1/34.5-/47-) for purging of occult breast cancer cells from bone marrow of patients. Here, we evaluate the therapeutic efficacy of this oncolytic virus. EXPERIMENTAL DESIGN: Electron microscopy was used to determine whether human breast cancer and bone marrow cells are permissive for JS1/34.5-/47- infection. Subsequently, the biological effects of JS1/34.5-/47- infection on human breast cancer cells and bone marrow were established using cell proliferation and colony formation assays, and the efficiency of cell kill was evaluated. Finally, the efficiency of JS1/34.5-/47- purging of breast cancer cells was examined in cocultures of breast cancer cells with bone marrow as well as bone marrow samples from high-risk breast cancer patients. RESULTS: We show effective killing of human breast cancer cell lines with the JS1/34.5-/47- virus. Furthermore, we show that treatment with JS1/34.5-/47- can significantly inhibit the growth of breast cancer cell lines without affecting cocultured mononuclear hematopoietic cells. Finally, we have found that the virus is effective in destroying disseminated tumors cells in bone marrow taken from breast cancer patients, without affecting the hematopoietic contents in these samples. CONCLUSION: Collectively, our data show that the JS1/34.5-/47- virus can selectively target breast cancer cells while sparing hematopoietic cells, suggesting that JS1/34.5-/47- can be used to purge contaminating breast cancer cells from human bone marrow in the setting of autologous hematopoietic cell transplantation.