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。     

Bone marrow micrometastases in breast cancer patients

We designed three new four–drug cisplatin–containing combinations and evaluated their activity in a randomized phase II study including patients with locally advanced (stage III) and locally recurrent breast carcinoma.All combinations included methotrexate (M) on day 1 and cisplatin (P) on day 2 (MVAC–like combinations) and differed from one another by the addition of Epirubicin (Epi), Vincristine (V), Etoposide (E), Mitomycin (Mi). Based on the administered agents, they were named MPEMi, MPEpiE, MPEpiV. The combinations were randomly assigned to 101 patients, 57 with locally advanced and 44 with locally recurrent breast carcinoma. Response was evaluated after 4 cycles.The complete response (CR) rates were 7 and 43 and the CR plus partial response (PR) rates were 84 and 89 in locally advanced and in locally recurrent disease, respectively. In locally advanced disease, a pathologic CR (pCR) was assessed in seven of 57 patients (12). There were no significant differences among the three combinations. The toxicities were at times severe, but generally tolerable, as demonstrated by the high cumulative doses of the drugs received by the patients.In conclusion, these three innovative chemotherapy regimens induced high CR plus PR rates in the neoadjuvant treatment of stage III and of locally recurrent breast carcinoma, and a high rate of pCR in stage III disease. These regimens warrant testing in phase III trials.     

The fate of bone marrow micrometastases in patients with primary breast cancer

Using an immunocytochemical technique, micrometastases have been found in the bone marrow of approximately 26% of patients with primary breast cancer at the time of initial surgery. To determine the fate of these cells, both in patients receiving and not receiving adjuvant therapy, multiple bone marrow aspirates were repeated in 82 patients at a median time of 18 months after surgery but prior to overt relapse. In both treated and untreated patients micrometastases were only found in one of 45 (2%) and one of 37 (3%) patients, respectively. However, when multiple marrow aspirates were taken from patients with local recurrence the incidence of micrometastases was 19% (three of 16), and this increased to 30% (three of ten) in patients with disease at distant sites other than bone, and 100% (ten of ten) in patients with radiologically proven bony disease. Three of 11 (27%) patients in whom the primary tumor remained in situ while receiving adjuvant therapy before definitive surgery had micrometastases at the time of diagnosis and at follow-up 3 months later. These results suggest that many of the micrometastases from breast cancer patients are the result of "shedding" of cells from the primary carcinoma and that a proportion are not viable. The technique is currently insufficiently sensitive to accurately monitor adjuvant therapy in breast cancer patients.     

Prognostic significance of micrometastases in bone marrow in patients with primary breast cancer

Metastatic breast cancer cells were found in the bone marrow of 60 (23%) of 269 patients with primary breast cancer, none of whom had metastatic disease disclosed by any other investigation, including bone scanning and radiological skeletal survey. We estimated the number of cancer cells as less than or more than 20 cancer cells seen. Twenty-six patients had less than 20 cancer cells present, and 34 had 20 or more. At a median follow-up time of 22 months, 53 patients had relapsed, 19 of 60 (31.7%) in the group found to have micrometastases and 34 of 195 (17.2%) in the group that had normal bone marrow. Patients with micrometastases are relapsing at a faster rate than those without micrometastases (P = less than 0.05). Patients with less than 20 cancer cells present are relapsing faster than those with no cancer cells but slower than those with 20 or more cancer cells (P = less than 0.01). We conclude that the presence of cancer cells in the marrow at primary diagnosis is a prognostic factor in patients with primary breast cancer.     

Detection of cytokeratin-positive cells in the bone marrow of breast cancer patients undergoing adjuvant therapy

Summary The presence of cytokeratin-positive cells in the bone marrow of breast cancer patients has been proven to be an independent prognostic factor. Their fate in primary breast cancer patients undergoing adjuvant therapy is of particular interest. We investigated the bone marrow status of 112 patients undergoing postoperative adjuvant treatment before and after therapy. A total of 373 patients with histologically confirmed primary breast cancer underwent bone marrow aspiration at the time of primary surgery. All patients were informed of their bone marrow status and offered repeat aspiration after 12 months. All patients were then treated with adjuvant chemotherapy, endocrine therapy or both based on current treatment recommendations. About 112 patients returned for a second bone marrow aspiration after a mean interval of 12 months following the initiation of adjuvant treatment. In 93 of 112 patients (83%) disseminated tumor cells had been found in the bone marrow before initiation of systemic chemo/endocrine therapy. At the time of follow-up sampling, after surgery and completion of adjuvant chemotherapy, the positivity rate dropped to 24%. Positive bone marrow status during follow-up was only associated with grading (p=0.020). Adjuvant treatment regimens are not able to completely eliminate cytokeratin-positive cells from the bone marrow. Prospective studies need to evaluate, whether these cells could become targets for additional adjuvant therapy.     

Comparative analysis of micrometastasis to the bone marrow and lymph nodes of node-negative breast cancer patients receiving no adjuvant therapy.

PURPOSE: In node-negative patients, of whom up to 30% will recur within 5 years after diagnosis, markers are still needed that identify patients at high enough risk to warrant further adjuvant treatment. In the present study we analyzed whether a correlation exists between microscopic tumor cell spread to bone marrow and to lymph nodes and attempted to determine which route is clinically more important. PATIENTS AND METHODS: According to a prospective design, bone marrow aspirates and axillary lymph nodes of level I (n = 1,590) from 150 node-negative patients with stage I or II breast cancer were analyzed immunocytochemically with monoclonal anticytokeratin (CK) antibodies. We investigated associations with prognostic factors and the effect of micrometastasis on patients' prognosis. RESULTS: CK-positive cells in bone marrow aspirates were present in 44 (29%) of 150 breast cancer patients, whereas only 13 patients (9%) had such positive findings in lymph nodes; simultaneous microdissemination to bone marrow and lymph nodes was seen in merely two patients. No correlation of bone marrow micrometastases with other risk factors was assessed. Reduced 4-year distant disease-free and overall survival were each associated with a positive bone marrow finding (P =.032 and P =.014, respectively) but not with lymph node micrometastasis. Multivariate analysis revealed an independent prognostic effect of bone marrow micrometastasis on survival, with a hazards ratio of 6.1 (95% confidence interval, 1.2 to 31.3) for cancer-related death (P =.031) in our series. CONCLUSION: Immunocytochemical detection of micrometastatic cells in bone marrow but not in lymph nodes is an independent prognostic risk factor in node-negative breast cancer that may have implications for surgery and stratification into adjuvant therapy trials.     

乳腺癌骨髓微转移的检测及意义

骨髓和骨是乳腺癌最易发生转移的部位。随着研究的不断深入，人们对骨髓微转移的检测和意义有了一定了解。大量研究结果表明，乳腺癌骨髓微转移与肿瘤预后、治疗、判断疗效有一定关系。现简介乳腺癌骨髓微转移的检测和意义。     

Comparative analyses of bone marrow micrometastases in breast and gastric cancer

This study is a comparative analysis of the prevalence, absolute number and aggregation status of bone marrow micrometastases (BMM) between breast (n = 234) and gastric (n = 102) cancer patients based on a standardized number of 1 × 10⁶ bone marrow-derived cells per patient. Additionally, expression of the epithelial cell adhesion molecule E-cadherin was analyzed on disseminated tumor cells. A positive BMM status was demonstrated in 88/234 breast and 45/102 gastric cancer patients. The presence of CK18⁺ cells positively correlated with parameters of advanced tumor progression in breast, but not in gastric cancer. Interestingly, 25.2% of the node-negative patients already had micrometastatic cells in the bone marrow at diagnosis. Regarding the absolute number of CK18⁺ cells and the frequency of CK18⁺ cell clusters, no significant difference was found between the 2 tumor types. However, clusters consisting of more than 10 CK18⁺ cells (type II clusters) were present exclusively in breast cancer patients. Additionally, co-expression of CK18 and E-cadherin was detectable in 15/21 micrometastases-positive breast but in only 1/9 gastric cancer patients. While prevalence of micrometastatic cells in bone marrow is discussed as an early indicator for systemic disease, aggregation status and a certain antigen profile might be indicative for site-specific differences in the manifestation pattern of solid metastases. © 1996 Wiley-Liss, Inc.     

Quality of life assessment in patients receiving adjuvant therapy for breast cancer: The IBCSG approach

Background and purpose: The International Breast Cancer Study Group (IBCSG) has developed an approach for assessing the impact of adjuvant therapy on quality of life (QL) within the framework of international, multilingual clinical trials. The major steps are summarized. Conceptual, methodological and practical issues are discussed with reference to results of two trials closed to accrual (IBCSG VI, VII) and one subsequent ongoing trial (IBCSG IX).Patients and methods: QL was assessed in pre- and postmenopausal patients with operable breast cancer. Various single-item linear analogue self-assessment (LASA) scales were used as indicators of components of QL, including global indicators of well-being, functioning and health perception, and specific indicators of symptoms of disease and treatment. In trials VI and VII, QL was assessed at baseline, during adjuvant treatment and follow-up, and at recurrence. Based on this experience, the QL form was revised for subsequent trials and further investigated in a subsample of patients randomized into trial IX.Results: In trials VI and VII, the QL indicators were responsive to the impact of biomedical factors at baseline, various adjuvant treatments, changes over the first 18 months, and recurrence. In trial IX, the revised QL form was well accepted by patients and staff. Completing this form did not exceed five minutes. QL differences between on and off cytotoxic treatment strengthen the claim that these measures are responsive. Correlations and logistic regression analyses show the expected relationship among the various global and specific indicators.Conclusion: Results from two trials closed to accrual and an ongoing trial confirm the feasibility, validity and clinical relevance of the IBCSG approach for studying the impact of adjuvant breast cancer therapy on QL in international clinical trials.     

Clinical significance of proliferative potential of occult metastatic cells in bone marrow of patients with breast cancer

There is increasing statistical evidence that the presence of tumour cells in bone marrow detected by immunocytochemistry represents an important prognostic indicator in breast cancer, but their individual capacity to become clinical metastases is unknown. The aim of this study was to assess the proliferative capacity of these occult metastatic cells in the bone marrow of patients with various stages of breast cancer. We obtained bone marrow aspirates from 60 patients with breast cancer before treatment with chemotherapy: 17 stage II, 12 stage III and 31 stage IV. After bone marrow culture for 6-34 days (median: 17 days) under specific cell culture conditions, viable epithelial cells were detected by cytokeratin staining in 40 patients (66%). Expansion of tumour cells was poorly correlated with tumour cell detection on primary screening (P=0.06). There was a nonsignificant correlation between the number and the presence of expanded tumour cells and the UICC stage of the patients. On primary screening, tumour cell detection was positive in 56% of patients and was correlated with clinical UICC stage (P=0.01). However, with a median follow-up of 23 months, expansion of tumour cells from bone marrow was associated with decreased patient survival (P=0.04), whereas the survival difference according to detection of CK-positive cells on primary screening was not statistically significant. In conclusion, viable tumour cells can be detected in the bone marrow of breast cancer patients. Their proliferative potential could be predictive of outcome and deserves further investigation.     

Axillary lymph node and bone marrow micrometastases of breast cancer]

Micrometastatic disease from breast cancer is a major concern both for clinicians and pathologists. Histologically, they can be defined as potentially invasive microfoci of tumoral cancer cells located in the vicinity of blood vessels and are a distinct entity from residual disease. They are mainly looked for in bone marrow and lymph nodes specimens and their diagnosis is currently easier thanks to immunohistochemistry. Provided a standard definition of micrometastatic disease and standard screening procedures can be met, the presence of micrometastases at the time of diagnosis could represent a major parameter in therapeutic decision-making. Although controversial, the presence of medullary and axillary lymph node micrometastases appears to be a major prognostic factor in terms of survival. Recognition of this entity could help in better defining the high-risk subset of patients who would potentially benefit from adjuvant chemotherapy.     

Immunocytochemical detection of epithelial cells in the bone marrow of primary breast cancer patients: a meta-analysis

Purpose. The immunocytochemical detection of epithelial cells in the bone marrow (BM) of breast cancer (BC) patients has been shown to have prognostic importance in several studies. We conducted a systematic review of the literature and a meta-analysis to assess the specificity, rate of positivity and correlation with known prognostic variables as well as with disease free (DFS) and overall survival (OS). Design. We performed a systematic review from the published literature. We included studies with at least 20 previously untreated BC patients as well as non-BC controls that evaluated the presence of epithelial cells in BM using immunocytochemistry (IC). Results. We identified 14 eligible studies with 3253 BC patients and 532 controls. Our results showed that IC has an overall rate of positivity of 31%(95%CI: 30–33%), and specificity of 96%(compared to normal patients) (95%CI: 0.94–0.98). We observed significant direct correlations between IC BM positivity and the primary tumor's presence with expression of estrogen receptors, larger size and higher histologic grade as well as with the presence of more than 3 positive axillary lymph nodes. At 5years of follow up IC positivity correlated directly with a lower DFS (relative risk=1.60; 95%CI: 1.39–1.83) and OS (relative risk=1.73; 95%CI: 1.29–2.31). The prognostic impact of BM positivity seemed to decrease with time. Conclusions. We conclude that the finding of epithelial cells in BM of BC patients correlates with several known prognostic factors and has adverse impact in the DFS and OS of these patients that seems, however, to decrease with time.     

Tattoo allergy in patients receiving adjuvant radiotherapy for breast cancer

Tattooing is routinely employed prior to radiotherapy treatment but allergies to tattoos are rare. New information on the incidence of tattoo allergy at St George Hospital is presented with details of two clinical cases. The literature on tattoo allergy has been unable to estimate the incidence of allergic reaction to tattoos because the total number of patients treated is unknown and not all patients were followed up. Our radiation oncology population for the first time has provided a known denominator, but wide confidence intervals prevent an accurate estimate of the incidence. Salient issues about tattoo allergy are highlighted based on a review of the published literature from 1966 to 1998.     

New strategies in marrow purging for breast cancer patients receiving high-dose chemotherapy with autologous bone marrow transplantation

Summary High-dose chemotherapy and autologous bone marrow transplantation (ABMT) are commonly used to treat selected patients with high-risk breast cancer. A limitation of ABMT is that clonogenic cancer cells could be collected with the bone marrow and produce a relapse of disease when reinfused into patients. Purging the marrowex vivo may eliminate the tumor cells, but it can also delay engraftment. We employed two different purging methods whereby breast cancer cells were depleted without delaying engraftment. The addition of WR-2721 (amifostine) to 4-hydroperoxycyclophosphamide (4-HC) reduced the time to engraftment by 10 days compared with marrow purged with 4-HC alone (26 versus 37 days, respectively). The positive selection of CD34+ hematopoietic progenitors produced engraftment within 21 days. The use of granulocyte colony-stimulating factor (G-CSF) accelerated the engraftment time of CD34+ hematopoietic progenitors to 11 days.Held in conjunction with the 15th Annual San Antonio Breast Cancer Symposium, December 1992, and supported by an educational grant from Lederle Oncology; editing by The Medicine Group USA, Inc.     

Final 5-year results of Z-FAST trial: adjuvant zoledronic acid maintains bone mass in postmenopausal breast cancer patients receiving letrozole

BACKGROUND:

Postmenopausal breast cancer (BC) patients receiving adjuvant aromatase inhibitor therapy are at risk of progressive bone loss and fractures. Zoledronic acid inhibits osteoclastic bone resorption, is effective in maintaining bone health, and may therefore be beneficial in this setting.

METHODS:

Overall, 602 postmenopausal women with early, hormone receptor‐positive BC receiving adjuvant letrozole were randomized (301 each group) to receive upfront or delayed‐start zoledronic acid (4 mg intravenously every 6 months) for 5 years. The primary endpoint was the change in lumbar spine (LS) bone mineral density (BMD) at month 12. Secondary endpoints included changes in LS BMD, total hip BMD, and bone turnover markers at 2, 3, and 5 years; fracture incidence at 3 years; and time to disease recurrence.

RESULTS:

At month 61, the adjusted mean difference in LS and total hip BMDs between the upfront and delayed groups was 8.9% and 6.7%, respectively (P < .0001, for both). Approximately 25% of delayed patients received zoledronic acid by month 61. Only 1 patient experienced grade 4 renal dysfunction; no confirmed cases of osteonecrosis of the jaw were reported. Fracture rates (upfront, 28 [9.3%]; delayed, 33 [11%]; P = .3803) and Kaplan‐Meier disease recurrence rates (upfront, 9.8 [95% confidence interval (CI), 6.0‐10.3]; delayed, 10.5 [95% CI, 6.6‐14.4]; P = .6283) were similar at month 61.

CONCLUSIONS:

Upfront zoledronic acid seems to be the preferred treatment strategy versus delayed administration, as it significantly and progressively increases BMD in postmenopausal women with early BC receiving letrozole for 5 years, and long‐term coadministration of letrozole and zoledronic acid is well tolerated. Cancer 2012. © 2011 American Cancer Society.     

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.     

Cyclin D1 expression in breast cancer patients receiving adjuvant tamoxifen-based therapy.

PURPOSE: The objective of our study was to determine the clinical relevance of cyclin D1 expression in hormone receptor-positive breast cancer patients who were treated with tamoxifen-based therapy. EXPERIMENTAL DESIGN: We assessed expression of cyclin D1 in surgical specimens of breast carcinoma by means of immunohistochemistry. Patients had been enrolled in either Austrian Breast and Colorectal Cancer Study Group (ABCSG) Trial 05 or ABCSG Trial 06 and received tamoxifen as part of their adjuvant treatment. Overall survival and relapse-free survival were analyzed with Cox models adjusted for clinical and pathologic factors. RESULTS: Cyclin D1 was expressed in 140 of 253 (55%) tumors of ABCSG Trial 05 and in 569 of 948 (60%) tumors of ABCSG Trial 06. Expression of cyclin D1 was associated with poor outcome in both cohorts. Overall survival was significantly shorter in patients with cyclin D1-positive tumors compared with patients with cyclin D1-negative tumors [adjusted hazard ratio (HR) for death (ABCSG Trial 05), 2.47; 95% confidence interval (95% CI), 1.08-5.63; P = 0.03; adjusted HR for death (ABCSG Trial 06), 1.78; 95% CI, 1.36-2.34; P < 0.0001]. Relapse-free survival was also shorter in patients with cyclin D1-positive tumors than in patients with cyclin D1-negative tumors [adjusted HR for relapse (ABCSG Trial 05), 2.73; 95% CI, 1.50-4.96; P = 0.001; adjusted HR for relapse (ABCSG Trial 06), 1.52; 95% CI, 1.14-2.04; P = 0.005]. CONCLUSION: Cyclin D1 expression is an independent poor prognostic factor in women with early-stage, hormone receptor-positive breast cancer who received adjuvant tamoxifen-based therapy.