小鼠动物模型中骨髓播散肿瘤细胞的检测方法

   探究小鼠骨髓中播散肿瘤细胞（disseminated tumor cells,DTCs ）的检测方法。   采用慢病毒感染的方法构建MDA-MB-231-GFP/Luc乳腺癌细胞系,将MDA-MB-231-GFP/Luc细胞经左心室接种于NOD-SCID小鼠体内,构建骨髓DTCs小鼠模型。采用荧光定量RT-qPCR、流式细胞计数、骨组织连续切片免疫荧光染色三种检测方法对小鼠骨髓DTCs进行定量和组织学定位研究。   荧光定量RT-qPCR法和流式细胞计数法的检测下限分别为22个和25个绿色荧光蛋白阳性（GFP⁺）细胞。骨组织连续切片免疫荧光染色法虽然不能定量骨髓DTCs数量,但可观察到GFP⁺ DTCs在骨组织中的分布,定位于成骨细胞或骨基质附近。   三种检测方法联合使用可满足小鼠骨转移研究动物实验中骨髓DTCs的定量和定位研究的需求,可为乳腺癌骨转移和骨髓中休眠癌细胞研究提供方法学支持。     

Detection of minimal residual disease in blood and bone marrow in early stage breast cancer

BACKGROUND:

The significance of circulating tumor cells (CTCs) in blood and of disseminated tumor cells (DTCs) in bone marrow (BM) in patients with early stage breast cancer is unclear. In this study, the authors investigated the occurrence of CTCs and DTCs in women with early stage breast cancer and evaluated the correlation of their presence with other prognostic markers.

METHODS:

Blood and BM aspirations were collected at the time of primary breast surgery. CTCs were detected by using the CellSearch assay, and DTCs were detected by immunostaining BM aspirates for pancytokeratin. The presence of CTCs and DTCs was correlated with tumor classification (T1 vs T2), tumor histologic grade, estrogen receptor (ER) status, progesterone receptor (PR) status, human epidermal growth factor receptor 2 (HER2) status, and lymph node (LN) status.

RESULTS:

Of 92 patients who were included in the study, 49 had T1 tumors, and 43 had T2 tumors. CTCs were detected in 31% of patients, and DTCs were detected in 27% of patients. There was no correlation between the occurrence of CTCs and DTCs with the tumor classification (T1 vs T2) or histologic grade. CTCs were detected in 33% of patients with ER‐positive disease versus 26% of patients with ER‐negative disease, in 32% of patients with PR‐positive disease versus 30% of patients with PR‐negative disease, and in 25% of patients with HER2‐positive disease versus 31% of patients with HER2‐negative disease. DTCs were observed in 23% of patients with ER‐positive disease versus 37% of patients with ER‐negative disease, in 22% of patients with PR‐positive disease versus 32% of patients with PR‐negative disease, and in 0% of patients with HER2‐positive disease versus 29% of patients with HER2‐negative disease. CTCs and DTCs were nearly equally prevalent in both LN‐positive women and LN‐negative women. There was no significant correlation between the occurrence of CTCs or DTCs with tumor classification (T1 vs T2), tumor histologic grade, positive ER status, positive PR status, or positive HER2 status, and axillary LN status.

CONCLUSIONS:

CTCs and DTCs in women with early stage breast cancer did not correlate with the standard prognostic indicators that were considered. The implications of their occurrence in patients with early stage disease will require further large‐scale studies. Cancer 2010. © 2010 American Cancer Society.     

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.     

Different clinical impact of estradiol receptor determination according to the analytical method: a study on 1940 breast cancer patients over a period of 16 consecutive years

SummaryBackground The persistence of disseminated tumor cells (DTC) in bone marrow (BM) of breast cancer patients is associated with poor prognosis. Preliminary studies indicated that these patients might benefit from secondary adjuvant targeted therapy. HER2 protein is suggested as one of the most promising targets. The aims of this study were (1) to determine the HER2 status of DTC in BM of breast cancer patients and (2) to compare the HER2 status of DTC and corresponding primary tumors.Methods BM aspirates from 137 primary breast cancer patients were included into the study. A double staining procedure was used for the identification of cytokeratin-positive (CK)/HER2 positive cells. HER2 status of the primary tumor was immunohistochemically assessed by the HERCEP-test™.Results In 46 of 137 (34%) breast cancer patients CK-positive cells were detectable in BM. DTC with HER2 positivity were found in 20 (43%) of these patients. The HER2 expression on DTC was heterogeneous in 7 of 17 (41%) patients. Concordance rate of HER2 status between primary tumor and DTC was 62%. In 12 of 20 patients with HER2 negative tumors HER2 positive DTC were detected.Conclusions HER2 positive DTC can be detected in patients with HER2 negative primary tumors. Therefore, the antigenic profile of DTC may be considered for treatment decision since these patients might actually benefit from trastuzumab. However, the HER2 overexpression on DTC is heterogeneous in individual patients which may reduce the efficacy of an immunotherapy based strategy directed against HER2-antigen only.     

骨髓坏死6例临床分析

目的：探讨骨髓坏死（BMN）的原发病、发病机理、临床特点及预后。方法：对6例骨髓坏死的临床资料进行分析。结果：6例BMN均以骨髓坏死为首要表现,以骨痛和发热为主要表现。1例原发病不明,其余5例均为恶性疾病,其中急性粒单核细胞白血病1例,急性淋巴细胞白血病2例,骨髓转移瘤2例。2例经积极治疗原发病取得一定疗效。结论：骨髓坏死是一种罕见的临床综合征,诊断难度大,预后差,应积极治疗原发病。     

骨髓坏死6例临床分析

目的:探讨骨髓坏死(BMN)的原发病、发病机理、临床特点及预后。方法:对6例骨髓坏死的临床资料进行分析。结果:6例BMN均以骨髓坏死为首要表现,以骨痛和发热为主要表现。1例原发病不明,其余5例均为恶性疾病,其中急性粒单核细胞白血病1例,急性淋巴细胞白血病2例,骨髓转移瘤2例。2例经积极治疗原发病取得一定疗效。结论:骨髓坏死是一种罕见的临床综合征,诊断难度大,预后差,应积极治疗原发病。     

骨髓转移癌的临床与实验室特征

本文对1984年至1991年间我院所见14例骨髓转移癌的临床及实验室特征进行了分析报道。发现本组女多于男(男:女=1:1.8).且女性发病年龄较轻。剧烈骨痛、高热、贫血、出血;骨压痛与骨髓有核细胞增生低下足常见的临床与实验室表现。所有病例经骨髓涂片和(或)骨髓活检均发现癌细胞。8例找到原发灶,其中4例为胃粘液腺癌,2例前列腺癌,另2例分别为肺癌及肝癌。本文对如何提高本病的确诊率进行了讨论。     

25例神经母细胞瘤临床报告

本文报告了25例神经母细胞瘤患儿的临床观察结果。其中,男性20例,女性5例。年龄最小为19个月,最大为8岁。5岁以下的患儿占72%(18/25)。23例(92%)患儿是经骨髓涂片检查确诊。由于本病原发肿瘤是多部位的且易早期广泛转移,症状的多样性是神经母细胞瘤最主要的临床特点之一。     

肿瘤预转移微环境形成的研究进展

摘 要：原发灶肿瘤通过诱导并改造未来转移器官,使之成为适合循环肿瘤细胞（circulatingtumor cells,CTCs）定植的微环境,即肿瘤预转移微环境（pre-metastatic niche,PMN）。PMN形成包括血管渗漏及通透性改变、凝血异常、细胞外基质重塑、骨髓源细胞的迁移和募集、免疫抑制等过程,其中肿瘤来源外泌体,肿瘤来源可溶性因子（tumor-derived soluble factors,TDSFs）,骨髓源细胞（bone marrow-derived cells,BMDCs）等在这过程中至关重要。全文结合国内外最新研究成果,对外泌体、TDSFs、BMDCs等在PMN形成过程中发挥的作用进行综述,探讨其涉及的具体机制以及可能发现针对肿瘤转移治疗的新靶点和可能面临的挑战。