乳腺肿瘤细胞TrfR表达与AgNOR计数的相互关系及临床应用价值

收集57例乳腺肿块穿刺或印片标本,用PAP法进行免疫化学染色观察TrfR阳性细胞和核仁组成区嗜银蛋白染色计算AgNORs颗粒数。结果提示:乳腺癌与非癌组织之间OKT_9,及AgNORs数的差别非常显著(P<0.01),而乳腺纤维腺瘤与腺病其差别无意义(P>0.05),OKT_9阳性与阴性乳腺癌之间AgNORs数的差别是显著的(P<0.05),呈正比关系。我们认为AgNORs和TrfR检测,对判断乳腺细胞其良恶性有十分重要意义,能作为细胞穿刺的辅助诊断手段。     

Silver-stained nucleolar organiser region counts are of no prognostic value in primary breast cancer.

The assessment of nucleolar organising regions have been reported to be of prognostic value both in a number of haematological and solid tumours. We have examined the relationship between the number of nucleolar organising regions (NORs) present in 75 primary breast cancers and various clinical and pathological features known to be associated with prognosis in patients with breast cancer. Formalin-fixed, paraffin-embedded tumour tissue was sectioned and stained by a one-stage argyrophil (AgNOR) method. Using light microscopy the mean number of AgNORs per cell was calculated. No correlation was observed between AgNOR counts and any of the prognostic variables studied, including oestrogen receptor (ER) status, histological grade of malignancy, lymph node stage or site of initial metastatic disease. Similarly there was no correlation between AgNOR counts and disease-free interval or survival. AgNOR counts do not appear to be a prognostic factor in primary breast cancer.     

Argyrophilic nucleolar organizer region (AgNOR) area per nucleus as a prognostic factor in breast cancer

Whether argyrophilic nucleolar organizer region (AgNOR) is a predictor of prognosis in breast cancer is controversial. The semiquantitative procedure used to calculate interphasic argyrophilic dots optically is neither reliable nor reproducible. We measured the mean area of AgNOR dots, the mean AgNOR area per nucleus (MA), and the mean AgNOR number per nucleus (MN) by automated image analysis in specimens from 131 patients with breast cancer. Higher MA (>7.41 pun²), detected in 22 (16.8%) of 131 patients, did not correlate with clinicobiologic variables, except for DNA ploidy status. Overall survival was significantly better in patients with lower MA than in those with higher MA. On multivariate analysis, MA was a significant independent factor, followed by nodal status. These findings indicate that expression of AgNOR should be evaluated in terms of area occupied and that the mean area of AgNOR per nucleus (MA) is an important prognostic factor in the overall survival of breast cancer patients. © 1995 Wiley-Liss, Inc.     

AgNOR技术在乳腺癌化疗药敏检测中的研究

目的：通过ＡｇＮＯＲ技术,针对不同的乳腺癌患者在四组联合化疗方案联合化疗方法,指导临床用药。方法：将５４例乳腺组织标本分别加入含有ＣＭＦ、ＣＡＤ、ＣＥＦ及ＥＴ联合化疗方案的培养基中进行培养,,应用ＡｇＮＯＲ技术检测出各组肿瘤细胞中嗜银蛋白含量,计算出各组联合化疗方案对被测患者的敏感顺序。结果：５４例乳腺癌２对所测的四组联合化疗方案反映出不同的敏感程度,各组间无显著差异,ＥＴ联合化疗方案对已而药的病     

Interphase nucleolar organizer region distribution in the tissues of oral leukoplakia, oral submucous fibrosis and oral cancer

The AgNOR technique, was applied to oral tissue sections of 185 oral cancer, 42 oral leukoplakia, 37 oral submucous fibrosis and 10 normal subjects to investigate whether any correlation held good in these different tissues. Compared to the AgNOR counts in normal oral epithelium, there was a gradation in increase in the mean AgNOR counts from oral leukoplakia to oral submucous fibrosis to oral carcinoma (P<0.01). This suggests that AgNOR count parallels with the degree of neoplastic transformation of oral epithelium. Three oral submucous fibrosis patients who showed very high AgNOR counts as that of oral cancer patients, later developed oral carcinoma. Among the oral cancer tissues, the moderately and poorly differentiated subtypes showed higher AgNOR counts and scattered distribution pattern than the well differentiated subtype which showed a clustered distribution pattern. These results suggest that AgNOR technique can be utilised as a diagnostic and prognostic indicator in premalignant and malignant oral tissues.     

Prevention of Mammary Carcinogenesis in C3H/OuJ Mice by Green Tea and Tamoxifen

Background: Tamoxifen (TAM) is useful in the chemoprevention of breast cancer, and green tea catechins,including (-)-epigallocatechin gallate (EGCG), may have similar actions. In this study, we investigated theireffects, alone or in combination, on mammary carcinogenesis using breast cancer cells and preneoplastic lesionsinC3H/OuJ mice. Methods: Growth inhibitory effects of EGCG and TAM on MCF-7 cells were evaluatedwith the anchorage-independent colony forming assay. The effects on mammary tumor carcinogenesis andpreneoplastic lesions were assessed in vivo using animals treated with GTE in drinking water (1%, 0.1%), or atamoxifen pellet (10 mg/ animal, subcutaneously inoculated) or both agents in combination (1%GTE + 10 mgTAM). The number and size of mammary tumors were measured weekly during treatment. At 48 weeks of age,mice were sacrificed for the examination of hyperplastic alveolar nodules (HAN) and argyrophilic nucleolarorganizer regions (AgNOR). Results: In the anchorage-independent growth assay, EGCG and TAM exhibiteddose-dependent antiproliferative effects on MCF-7 cells. In the tumor formation assay, tumor incidences weredecreased in the GTE, TAM, and GTE+TAM groups, particularly in the latter, in which no tumors developed.AgNOR counts were also significantly lower in the 1%GTE+TAM compared with the 1%GTE group, suggestingan additional anticarcinogenic effect. Conclusion: These data suggest that GTE and TAM, individually and incombination, have potential for chemoprevention of breast cancer.     

Influence of Neoadjuvant Chemotherapy on HER2/neu Status in Invasive Breast Cancer

IntroductionReliably estimating HER2/neu expression in breast cancer is important for predicting patient prognosis and optimizing adjuvant therapeutic strategies. In this retrospective cohort study, effects of NAC on HER2/neu status in invasive breast cancer were evaluated, and the related factors were analyzed.Patients and MethodsOne hundred thirty-one patients with primary breast cancer were treated with anthracycline- and/or taxane-based NAC. HER2/neu status was evaluated by IHC on core needle biopsies of primary tumors before NAC and surgical resection specimens of post-NAC residual breast cancers or tumor-positive axillary lymph nodes. Thirty-two pairs of specimens with discordant HER2/neu IHC scores were analyzed by fluorescence in situ hybridization (FISH).ResultsA significant difference in HER2/neu status by IHC between core needle biopsies and surgical resection specimens in patients receiving NAC was observed. After NAC, 23.4% (29 of 124) of tumors showed downregulated HER2/neu expression by IHC. Alterations of HER2/neu IHC scores did not significantly correlate with tumor subtype, pathologic response to NAC, adjuvant regimen, or time interval from the last chemotherapy to surgery. HER2/neu protein overexpression level was associated with favorable pathologic response to anthracycline and taxane-based chemotherapy. However, tumors with altered HER2/neu IHC scores after NAC revealed stable HER2/neu gene amplification/nonamplification by FISH analysis.ConclusionNeoadjuvant chemotherapy for breast carcinoma resulted in the HER2/neu status alteration by IHC, but they have stable gene amplification status by FISH. HER2/neu protein overexpression indicated greater sensitivity to neoadjuvant anthracycline- and taxane-based chemotherapy. Thus, retesting HER2/neu IHC status in residual tumors after NAC should be considered in order to optimize adjuvant systemic therapy.     

DNA methylation of polycomb group target genes in cores taken from breast cancer centre and periphery

We previously demonstrated that methylation of neugogenic differentiation 1 (NEUROD1) gene, a polycomb group target (PCGT) gene is a predictor of response to neoadjuvant chemotherapy in breast cancer. Here, we address the question whether NEUROD1 methylation provides clinical information independent from its expression level, and whether PCGT methylation is homogeneous in breast cancer. We examined: (1) NEUROD1 methylation and mRNA expression in 9 breast cancer cell lines and 63 tumour specimens, (2) DNA methylation in a training set of 55 PCGT genes taken from the centre (TUC) and periphery (TUP) of 15 breast cancer specimens, and compared this with 22 non neoplastic controls, and finally, (3) validated statistically significant genes in an independent set of 20 cases versus 18 controls. 8/9 cell lines demonstrated NEUROD1 methylation, whereas, there was only one cell-line that showed NEUROD1 expression. There was no association between methylation and expression in breast tumour specimens, with only 14% exhibiting NEUROD1 expression. Of the 55 PCGT genes analysed, 24% (13/55) were shown to be cancer specific (p < 0.05) with a receiver-operating-characteristic (ROC) area-under-the-curve (AUC) of >0.7 (range 0.71–0.95). DNA methylation accurately predicted the presence of cancer in both TUC and TUP. DNA methylation of PCGT genes predicts the presence of breast cancer and is not subject to tumour heterogeneity. Further work will reveal if methylation of PCGT genes will serve as a robust means for the clinical detection and assessment of breast cancer.     

The relationship of the neo-angiogenic marker, endoglin, with response to neoadjuvant chemotherapy in breast cancer

Endoglin (CD105) is upregulated in endothelial cells of tissues undergoing neovascularisation. A greater number of CD105-positive vessels predicts poor survival in breast cancer. We examine whether CD105 expression predicts response to neoadjuvant chemotherapy. Fifty-seven women (median age 50 years, range 29-70) received neoadjuvant chemotherapy for operable breast cancer. Immunohistochemical staining using monoclonal antibodies to CD105 and CD34 was performed on pretreatment biopsies and post-treatment surgical specimens. Individual microvessels were counted in 10 random fields at x 200 magnification. Median counts were correlated with clinical and pathological response using the Mann-Whitney U-test. Forty-five out of fifty-seven patients (79%) responded clinically, 22 (39%) responded pathologically. On pretreatment biopsies, clinical responders had significantly lower median CD105-positive vessel counts than nonresponders (median counts 5 and 9.3/high-power field (hpf), median difference=4.0/hpf, 95% CI 0.5-8.0/hpf, P=0.02). For pathological responders and nonresponders, median counts were 4.8 and 5.5/hpf (median difference -0.5/hpf, 95% CI=-2.5-2.0/hpf, P=0.77). CD34 expression (total microvessel density) did not correlate with response. Pretreatment CD105 expression predicts for clinical response to chemotherapy, with a lower initial count being favourable. Patients with high baseline new vessel counts or increased counts after conventional therapy may benefit from additional antiangiogenic therapy.     

Determining circulating endothelial cells using CellSearch system during preoperative systemic chemotherapy in breast cancer patients

Background

Circulating endothelial cells (CECs) have been studied as a biomarker for tumour progression and monitoring therapeutic effects. The CellSearch system is a semi-automated system that allows standardised analysis of CECs. This study assessed the clinical implications of CECs determined by the CellSearch system in breast cancer patients.

Methods

Seventy-six consecutive breast cancer patients (53 operable and 23 metastatic or recurrent) were enrolled for the study. Thirty-five patients with operable breast cancer received preoperative chemotherapy with a regimen based on anthracycline and/or taxane. CECs are defined as CD146⁺CD105⁺CD45⁻DAPI⁺ cells in the system. CD34 expression was examined using the additional channel in the system.

Results

A majority (4539 of 5183 cells, 88%) of CECs from patients with operable breast cancer were CD34-positive. Triple-negative cancers showed higher baseline CEC and CD34⁺CEC counts than the other types (P = 0.0387 and 0.0377, respectively). Low baseline CEC and CD34⁺CEC counts, and a low CD34 positive rate were associated with pathological complete response (pCR) of preoperative chemotherapy in patients with primary breast cancer (P = 0.046, 0.027 and 0.01, respectively). In multivariate analyses, the CD34 positive rate was significant for pCR (P = 0.021). During preoperative chemotherapy, CEC and CD34⁺CEC counts before each cycle of chemotherapy increased with taxane-based regimens (P = 0.0018 and 0.0008, respectively) but not with anthracycline-based regimens.

Conclusions

Baseline CEC, in particular CD34⁺CEC, counts and the CD34 positive rate might be useful for the prediction of treatment response of preoperative chemotherapy in patients with operable breast cancer.     

Can CD44+/CD24- Tumor Cells Be Used to Determine the Extent of Breast Cancer Invasion Following Neoadjuvant Chemotherapy?

Purpose

To investigate the distribution of CD44⁺/CD24^- cells in breast cancers in relation to tumor size before and after the administration of neoadjuvant chemotherapy.

Methods

CD44⁺/CD24^- tumor cells obtained from breast cancer specimens were characterized in vivo and in vitro using tumor formation assays and mammosphere generation assays, respectively. The distribution of CD44+/CD24- tumor cells in 78 breast cancer specimens following administration of neoadjuvant chemotherapy was also evaluated using immunofluorescence assays, and this distribution was compared with the extent of tumor invasion predicted by Response Evaluation Criteria in Solid Tumours (RECIST).

Results

In 27/78 cases, complete remission (CR) was identified using RECIST. However, 18 of these CR cases were associated with a scattered distribution of tumor stem cells in the outline of the original tumor prior to neoadjuvant chemotherapy. After neoadjuvant chemotherapy, 24 cases involved cancer cells that were confined to the tumor outline, and 21 cases had tumor cells or tumor stem cells overlapping the tumor outline. In addition, there were 6 patients who were insensitive to chemotherapy, and in these cases, both cancer cells and stem cells were detected outside the contours of the tumor volume imaged prior to chemotherapy.

Conclusion

CD44+/CD24- tumor cells may be an additional parameter to evaluate when determining the extent of breast cancer invasion.     

Ki-67 as a Predictor of Response to Neoadjuvant Chemotherapy in Breast Cancer Patients

Purpose

The objectives of this study were to assess the potential value of Ki-67 in predicting response to neoadjuvant chemotherapy in breast cancer patients and to suggest a reasonable cutoff value for classifying Ki-67 expression.

Methods

This study included 74 breast cancer patients who underwent surgery after anthracycline-based neoadjuvant chemotherapy between 2007 and 2012. We analyzed the clinical and immunohistochemical characteristics using core biopsy specimens obtained before neoadjuvant chemotherapy to determine their correlations with the response to chemotherapy.

Results

A clinical complete response was observed in 6 patients (8.1%); a clinical partial response, in 44 patients (59.5%); and clinical stable disease, in 24 patients (32.4%). A pathologic complete response (pCR) was observed in 10 patients (13.5%). In univariate analysis, estrogen receptor (ER) negativity (p=0.031), human epidermal growth factor receptor 2 (HER2) positivity (p=0.040), and high Ki-67 expression (p=0.036) were predictive factors for a pCR. In multivariate analysis, Ki-67 was the only independent predictor of a pCR (p=0.049). The analysis of Ki-67 values revealed that 25% was a reasonable cutoff value for predicting the response to chemotherapy. In subgroup analysis, a higher Ki-67 value (≥25%) was a significant predictive factor for the response to neoadjuvant chemotherapy, especially in ER-negative and HER2-positive breast cancer patients.

Conclusion

Ki-67 expression in breast cancer tissue may be an effective factor for predicting the response to neoadjuvant chemotherapy. We suggest that a 25% level of Ki-67 expression is a reasonable cutoff value for predicting a response to chemotherapy. Moreover, Ki-67 is a useful predictive factor for pCR, especially in patients with ER-negative and HER2-positive breast cancer.     

AgNOR在乳腺癌细胞核中的形态及定量研究

 本文对40例乳腺癌细胞核中的A_gNOR颗粒数目、大小、结构和形状进行了定量研究，并以40例乳腺良性病变做对照，在大颗粒、小颗粒和混合颗粒三项定量指标中，乳腺癌均明显高于乳腺良性病变（P<0.01），小颗粒定量特异度和灵敏度为100%。乳腺癌组织类型不同、恶性程度不同。A_gNOR数目、结构和形状也有不同。作者认为准确合理划分A_gNOR颗粒大小，用多参数做定量指标，结合颗粒结构和形状分析，对乳腺癌诊断有重要价值，对协助判断乳腺癌恶性程度及预后也有重要意义。     

Bax expression has prognostic significance that is enhanced when combined with AgNOR counts in glottic carcinomas.

Using nucleolar organizer regions (NORs) as a proliferative marker and Bax expression as a marker for apoptosis, we have studied the individual and combined prognostic significance of these markers. Successive sections of diagnostic, formalin-fixed and paraffin-embedded specimens from 69 patients with T1-4 tumours were stained with a rabbit anti-human Bax polyclonal antibody and silver nitrate for visualization of NORs (AgNORs). After classification for staining intensity and the percentage of Bax expression, a final score resulting in four classes of increasing Bax expression was obtained. AgNOR counts were expressed as mean counts (mAgNOR) and the percentage of tumour nuclei with more than one AgNOR (pAgNOR>1). Both AgNOR parameters were grouped in three classes with increasing values. Low Bax scores correlated significantly with poor prognosis (P = 0.0106). For mAgNOR and pAgNOR>1, high values correlated with poor prognosis (P = 0.0185 and P = 0.0003 respectively). A combined parameter, for which the Bax score was subtracted from the AgNOR scores, appeared to be statistically stronger than the individual parameters (P < 0.0001). Both Bax expression and AgNOR scores, and in particular the combination of these parameters, appear to be strong prognostic markers in glottic squamous cell carcinomas.     

Evaluation of thyroid cancer in Chinese females with breast cancer by vascular endothelial growth factor (VEGF), microvessel density,and contrast-enhanced ultrasound (CEUS)

To evaluate thyroid cancer in Chinese females with breast cancer by VEGF, MVD, and contrast-enhanced ultrasound (CEUS), 34 of 2,278 female inpatients with breast diseases who underwent routine thyroid ultrasonography were pathologically proved as thyroid cancer and enrolled into two groups: a breast cancer group and a non-breast cancer group. CEUS was performed and enhancement patterns were classified. Time-intensity curve parameters were analyzed to correlate with MVD CD34 and VEGF expression. Fourteen (2.6 %) and 20 (1.1 %) patients in breast cancer and non-breast cancer group were pathologically diagnosed as thyroid cancer. Six (42.8 %) and 0(0 %) patients showed high enhancement CEUS patterns of thyroid cancer in these two groups, respectively. The arrival time of time-intense curve was shorter, and the mean and peak intensity were higher in thyroid cancer in breast cancer group. The mean MVD counts and VEGF expression were significantly higher in thyroid and breast carcinomas in breast cancer group (P?<?0.01). We also found that the mean and peak intensity were significantly associated with MVD counts and VEGF expression (P?<?0.01). CEUS is recommended in evaluating the microcirculation of thyroid cancer in women with breast cancer and has the significant relationship with MVD counts and VEGF expression.     

A central review of histopathology reports after breast cancer neoadjuvant chemotherapy in the neo-tango trial

Background:

Neo-tAnGo, a National Cancer Research Network (NCRN) multicentre randomised neoadjuvant chemotherapy trial in early breast cancer, enroled 831 patients in the United Kingdom. We report a central review of post-chemotherapy histopathology reports on the surgical specimens, to assess the presence and degree of response.

Methods:

A central independent two-reader review (EP and HME) of histopathology reports from post-treatment surgical specimens was performed. The quality and completeness of pathology reporting across all centres was assessed. The reviews included pathological response to chemotherapy (pathological complete response (pCR); minimal residual disease (MRD); and lesser degrees of response), laterality, the number of axillary metastases and axillary nodes, and the type of surgery. A consensus was reached after discussion.

Results:

In all, 825 surgical reports from 816 patients were available for review. Out of 4125 data items there were 347 discrepant results (8.4% of classifications), which involved 281 patients. These involved grading of breast response (169 but only 9 involving pCR vs MRD); laterality (6); presence of axillary metastasis (35); lymph node counts (108); and type of axillary surgery (29). Excluding cases with pCR, only 45% of reports included any comment regarding response in the breast and 30% in the axillary lymph nodes.

Conclusion:

We found considerable variability in the completeness of reporting of surgical specimens within this national neoadjuvant breast cancer trial. This highlights the need for consensus guidelines among trial groups on histopathology reporting, and the participation of histopathologists throughout the development and analysis of neoadjuvant trials.     

Argyrophilic nucleolar organiser region counts and prognosis in pharyngeal carcinoma.

The prognostic significance of argyrophilic nucleolar organiser regions (AgNORs) has been evaluated in biopsy specimens from 61 primary squamous and undifferentiated carcinomas of the pharynx prior to therapy. The univariate Kaplan-Meyer survival analysis showed a significant correlation between 3- and 5-year survival rates and the mean AgNOR number per tumour cell (P less than 0.001). No significant correlation was found between prognosis and patients age and sex, tumour location, clinical stage, histologic grade, extent of lymphocytic infiltration, HMFG-2 positivity of tumour cells and UCHL1, LN2, MB2 positivity of infiltrating lymphocytes. There was no significant association between AgNOR counts and tumour histologic grade or clinical stage. Multivariate survival analysis showed that only two variables were significantly correlated with prognosis: AgNOR counts (P less than 0.001) and the extent of lymphocytic infiltration (P less than 0.027). Our results indicate the prognostic value of AgNOR counts and suggest the use of this method as a significant parameter in the pretherapeutic assessment of the aggressiveness of pharyngeal carcinomas.     

Impact of Oncotype DX Recurrence Score in the Management of Breast Cancer Cases

BackgroundOncologists have used clinicopathologic features to guide treatment decisions for their breast cancer patients; however, more recently, results of multigene assays are also being considered. A popular assay, Oncotype DX (Genomic Health), stratifies node-negative breast cancer patients into groups that are at low, intermediate, or high risk for distant recurrence and guides decisions about adjuvant chemotherapy utilization.ObjectiveWe studied the impact of Oncotype DX recurrence score (ODxRS) compared with that of clinicopathologic features on adjuvant chemotherapy utilization in node-negative breast cancer patients and in node-positive breast cancer patients, and we evaluated whether clinicopathologic features impact the decision for adjuvant chemotherapy utilization in a subset of node-negative breast cancer patients with an intermediate-risk ODxRS.MethodsA retrospective study from a single academic institution was performed on 425 patients with invasive breast carcinoma.ResultsAdjuvant chemotherapy utilization most significantly correlated with a high-risk ODxRS (P < .0001) and, to a lesser degree, patient's age and tumor size. No statistically significant association was found between ODxRS and adjuvant chemotherapy utilization in a subset of patients. In the 156 node-negative breast cancer patients with intermediate-risk ODxRS, high tumor grade most significantly correlated with adjuvant chemotherapy utilization (P < .0001).ConclusionODxRS, if available, heavily impacts adjuvant chemotherapy utilization and more so than any clinicopathologic factor in node-negative breast cancer patients. Node-negative breast cancer patients in the intermediate-risk group whose tumors were high grade were more likely to receive adjuvant chemotherapy.