乳腺导管不同类型增生性病变中p63和CK5/6的表达及意义

目的：明确不同类型的乳腺导管增生性病变中p63和Cytokeratin 5/6(CK5/6)的表达和意义。方法：对150例乳腺导管增生性病变中的368个病灶分别进行p63和CK5/6染色。结果：CK5/6在正常乳腺组织中的肌上皮和导管上皮均阳性表达,35例普通型增生中,33例CK5/6呈马赛克式阳性表达,仅2例弥漫阳性;在非典型性增生、原位癌和浸润癌中,CK5/6主要呈阴性表达,仅6例阳性,其中有2例雌激素受体（estrogen receptor, ER）、孕激素受体（progesterone receptor, PR）和CerbB 2阴性,符合基底细胞样癌的特点。p63在良性病变和非浸润性肿瘤性增生病变中的肌上皮细胞阳性表达,3例原位癌不表达p63和CK5/6。浸润癌中有3例（3/43）p63散在肿瘤细胞阳性,其余均阴性。结论：不同类型乳腺增生性病变中p63及CK5/6的染色模式存在一定规律,有助于这些病变的诊断和鉴别诊断。     

乳腺增生性病变中p63蛋白的表达及意义

目的 探讨p63蛋白在乳腺疾病诊断以及鉴别诊断中的应用价值。方法 对38例不同类型的乳腺病变分别进行p63、SMA、S-100蛋白、CK(34βE12)免疫标记。结果 p63蛋白在乳腺良性增生性病变腺体肌上皮细胞核有阳性表达,围绕在腺体周围,原位癌癌巢周围肌上皮细胞数目减少,呈间断性分布,浸润性癌癌巢周围肌上皮细胞大部分消失或无肌上皮细胞分布,且阳性表达强度与肿瘤分化程度相关。其他基底细胞标记物也可以比较特异地显示乳腺腺体周围肌上皮,但存在一定的非特异性着色。结论 从乳腺正常组织、良性病变至原位癌、早期浸润和浸润性癌,腺体周围的肌上皮细胞呈逐渐消失的趋势,p63蛋白标记可以比较特异地反映这个过程,而且受其他影响因素小。     

Impact of immunohistochemical markers, CK5/6 and E-cadherin on diagnostic agreement in non-invasive proliferative breast lesions

Aims:  To assess the impact of cytokeratin (CK) 5/6 and E-cadherin immunohistochemistry on diagnostic agreement of non-invasive proliferative breast lesions.
Methods and results:  Twenty pathologists classified 105 cases of non-invasive proliferative breast lesions into 10 diagnostic categories. One haematoxylin and eosin (H&E) slide of each case was analysed on a first round and one H&E slide with corresponding CK5/6 and E-cadherin immunohistochemistry was analysed on a second round. Interobserver reproducibility for category-specific and management-specific lesions was measured on each round. CK5/6 and E-cadherin had little impact on diagnostic agreement, which remained moderate between the first and second rounds (overall κ coefficients of 0.47 and 0.53, respectively, P  = NS). Levels of agreement slightly improved for lesions with specific CK5/6 and E-cadherin immunoprofiles (usual ductal hyperplasia, atypical ductal hyperplasia, atypical lobular hyperplasia, lobular carcinoma in situ , non-high-grade ductal carcinoma in situ ), but the differences observed were not statistically significant. However, diagnostic agreement improved when lesions were grouped according to their management category (overall κ coefficients of 0.58 and 0.66 in the first and second rounds, respectively).
Conclusions:  CK5/6 and E-cadherin immunohistochemistry has little impact on interobserver reproducibility for non-invasive breast lesions. Diagnostic agreement can, however, be improved by grouping lesions in management categories.     

The significance of combined CK5/6 and p63 immunohistochemistry in predicting the risks of subsequent carcinoma development in intraductal papilloma of the breast

Prediction of subsequent risks of breast carcinoma (BC) development in intraductal papilloma (IDP) has remained controversial with the exception of atypical papilloma (AP). The potential value of immunohistochemistry (IHC) of cytokeratin 5/6 [CK5/6] and p63 have been proposed but its standardization has also remained controversial. We studied 17 patients initially diagnosed as IDP or AP who subsequently developed BC with 34 age‐matched controls. We compared histological features, results of IHC (estrogen receptor [ER], progesterone receptor [PR], human epidermal growth factor receptor 2 [HER2], p63, CK5/6, Ki67), and ultrasound findings. Univariate conditional logistic regression analysis revealed that the status of both CK5/6 and p63/CK5/6 were significantly associated with subsequent BC development (P < 0.05). BC development in CK5/6 positive patients was 17.9% and p63/CK5/6 double positive patients 8.6%, respectively. Ultrasound evaluation was not significantly associated with any of the parameters examined and subsequent carcinoma development. Despite CK5/6 positivity, the subsequent incidence of BC development was nearly 20%. However p63/CK5/6 double positive status could predict a significantly lower subsequent carcinoma incidence, indicating a more accurate prognostic utility. Combining p63/CK5/6 with histological findings could be easily applied and could predict the subsequent BC development of the patients diagnosed as IDP at biopsy.     

The significance of the diagnosis of atypia in breast fine-needle aspiration

The diagnosis of atypia in breast fine-needle aspiration (FNA) continues to be an area of debate in cytology practice. The aim of this study was to assess the clinical significance of this term and to evaluate potential morphological criteria, which would determine the patient's outcome. A computer-based search was carried out to retrieve breast FNAs performed between 1990 and 2000 that were diagnosed as atypical. Cases followed by surgical resection were reexamined for the presence of morphological features potentially differentiating benign and malignant lesions. Out of 1,568 breast FNAs, there were 64 cases (4%) with a diagnosis of atypia. Thirty-eight cases had surgical follow-up material that revealed malignancy in 14 cases (37%) and benign lesions in 24 cases (63%). The benign diagnostic categories included fibrocystic change (12/24), fibroadenoma (3/24), tubular adenoma (2/24), and nonspecific findings (7/24). The malignant diagnoses included ductal carcinoma (9/14), lobular carcinoma (3/14), ductal carcinoma in situ (DCIS; 1/14), and tubular carcinoma (1/14). The evaluation of cytological criteria used to differentiate benign from malignant lesions (i.e., cellularity, loss of cohesion, myoepithelial cells, nuclear enlargement, nuclear overlap, prominent nucleoli) revealed significant overlap between benign and malignant cases, particularly in cases of fibroadenoma, tubular adenoma, and proliferative breast disease. The surgical follow-up of four hypocellular cases revealed lobular carcinoma in two cases and ductal carcinoma in the remaining two cases. Our study confirmed that the diagnosis of atypia is clinically significant because it is associated with a high probability of malignancy. No morphological criterion is able to reliably differentiate benign and malignant lesions in cases diagnosed with atypia. Diagnosis of atypia is particularly significant in hypocellular cases. We recommended that breast FNAs with a diagnosis of atypia be evaluated further histologically.     

Expression of p63, keratin 5/6, keratin 7, and surfactant-A in non-small cell lung carcinomas

In this study, we sought to validate the importance of p63, CK5/CK6, CK7, and surfactant-A (SP-A) to classify 42 non-small cell lung cancers in autopsy and surgical resection specimens and to study the usefulness of these markers in distinguishing between squamous cell carcinomas and adenocarcinomas because of their different implications regarding treatment and prognosis. All adenocarcinoma cases were negative for p63; 9 (56.2%) of 16 were CK5/CK6 positive, 16 (94.1%) of 17 were CK7 positive, and 4 (26.6%) of 15 were SP-A positive. In squamous cell carcinoma, 1 case was CK7 and SP-A positive and 14 (77.8%) of 18 were p63 positive. The latter appears to be useful in differentiating squamous cell carcinoma from adenocarcinoma of the lung in small biopsies without keratinization or glandular differentiation; thus, for advanced-stage cases, where there is no possibility of surgical resection and the treatment of choice is radiotherapy plus chemotherapy, we would be able to differentiate between the two histological types, establishing then a different therapy.     

CK5/6在乳腺浸润性小叶癌中的表达及意义

目的 研究CK5/6在乳腺浸润性小叶癌(invasive lobular carcinoma,ILC)中的表达及其意义.方法 用免疫组化MaxVision方法检测61例ILC组织中CK5/6、ER、PR、Her-2及Ki-67等的表达情况.结果 61例ILC中,6例表达CK5/6,其中弥漫阳性3例,灶状阳性3例;组织学类型为混合型、多形型或实性型,核级为2～3级;5例核分裂小于5个/10 HPF,1例约20个/10 HPF;其中2例表达ER和PR,4例不表达;6例Her-2均阴性;3例Ki-67增殖指数大于60%.结论 ILC中有少数表达CK5/6,倾向于高核级、ER、PR、Her-2表达阴性;可能存在基底细胞样亚型小叶癌.     

718例浸润性乳腺癌分子分型中CK5/6和EGFR的表达及临床病理分析

目的 探讨CK5/6和EGFR在浸润性乳腺癌(invasive breast carcinoma,IBC)分子分型中的表达及相关性.方法 采用免疫组化法检测718例IBC中CK5/6、EGFR、ER、PR、HER-2及Ki-67蛋白的表达,分析CK5/6、EGFR与IBC组织学类型、分子分型及其他相关标志物之间的关系....     

p63显示肌上皮细胞在乳腺癌诊断中价值

目的比较p63与S-100蛋白、CK(34βE12)、CKl7、actin及calpornin在乳腺上皮良恶性病变中阳性表达的差异，显示肌上皮细胞在乳腺癌诊断中的价值。方法 筛选出含有正常乳腺组织及导管上皮不典型增生的导管原位癌6例，浸润型导管癌20例。根据《诊断外科病理学》标准将这些病变分为5组。选用p63等6种免疫组化抗体对上述病变进行标记。结果 (1)6种抗体在浸润癌以外的各种乳腺导管上皮增生性病变中均可见肌上皮细胞的阳性表达，在浸润癌中很少能找到肌上皮细胞．而在原位癌的癌巢周可见到完整或不连续的肌上皮细胞。(2)p63、actin及calponin在除浸润癌外的四组病变中均可见染色阳性的肌上皮细胞，阳性率达100％。但actin及calponin也可使血管平滑肌及肌纤维母细胞着色。(3)CKl7在正常及增生病变中肌上皮细胞阳性率明显低于不典型增生及原位癌。(4)S-100蛋白及CK(34βEl2)在各组肌上皮细胞染色阳性率较低,并可见分泌上皮阳性着色。结论 通过5组不同病变的观察，MEC的存在与否不能成为导管原位癌与不典型增生的鉴别标准,但MEC的有无却是乳腺原位癌与浸润癌的鉴别点之一。p63对乳腺肌上皮细胞最敏感，是鉴别乳腺原位癌与浸润癌的首选抗体。actin与calponin亦是乳腺肌上皮细胞最敏感的抗体，在判断乳腺癌有无浸润时,同时选用胞核阳性的p63及胞质阳性actin或calponin，是一种较科学的办法。     

Utility of WT-1, p63, MOC31, mesothelin, and cytokeratin (K903 and CK5/6) immunostains in differentiating adenocarcinoma, squamous cell carcinoma, and malignant mesothelioma in effusions

To distinguish carcinoma, either adenocarcinoma (ADC) or squamous cell carcinoma (SCC), and malignant mesothelioma (MM) in effusion can be a diagnostic challenge based on morphology alone. This study evaluates the utility of WT-1, p63, MOC31, mesothelin, and cytokeratin (K903 and CK5/6) immunostains in effusions when ADC and SCC of the lung are in the differential diagnosis with MM. A cohort of 43 effusions consisting of lung ADC (N = 10), SCC (N = 15), and MM (N = 18, mostly (16) pleural based), was subjected to immunostains using the above mentioned antibodies. WT-1 was positive in 100% MM, 0% ADC, and 0% SCC cases while p63 was positive in 0% MM, 30% ADC, and 80% SCC cases. Stain for MOC31 was positive in 100% ADC, 67% SCC, and 35% MM cases. Similarly, mesothelin antibody stained 100% ADC, 60% SCC, and 47% MM cases. Antibodies for K903 and CK5/6 stained 100% SCC cases but fewer ADC cases (40 and 10%, respectively). In conclusion, in this cohort of mostly pleural malignant effusion, MM can be identified with positive staining for WT-1 and negative staining for p63. Conversely, negative staining with WT-1 and positive staining for p63 exclude MM. Used as part of an immunostain panel, cytokeratin markers (CK5/6 and K903) are useful in differentiating SCC from ADC when MM is already excluded, and MOC31 might have limited value in differentiating ADC from MM. A negative stain with MOC31 can exclude lung ADC. Mesothelin, on the other hand, is not useful in the differential diagnosis of ADC, SCC, and MM.     

Clinical significance of CK5/6 and PTEN protein expression in patients with bilateral breast carcinoma

AIMS: To assess the expression of cytokeratin (CK) 5/6 in bilateral breast cancers and to assess the relationship between its expression and other prognostic variables, as well as between CK5/6 expression and patients' survival. METHODS AND RESULTS: The expression of CK5/6, PTEN protein, oestrogen receptor, progesterone receptor, p53 and c-erbB-2 protein were evaluated by immunohistochemistry in 88 primary breast cancers diagnosed in 44 women. To assess the prognostic value of studied factors, Cox regression analysis was performed. Expression of CK5/6 was found in 23 of 88 primary breast carcinomas (23/88; 26%). The hazard ratio of development of distant metastasis in patients in whom at least one cancer was CK5/6+ was 99.8 (P=0.037) and in patients with at least one carcinoma with reduced PTEN expression it was 10.8 (P=0.044). CK5/6 expression was correlated with absence of oestrogen (P<0.0001) and progesterone receptors (P<0.0001) and very strong expression of p53 (P<0.05). Reduced PTEN expression was correlated with presence of axillary metastases (P<0.01), with very strong expression of c-erbB-2 (P<0.05) and with reduced expression of oestrogen receptor (P<0.05). CONCLUSIONS: Analysis of expression of CK5/6 and PTEN protein in bilateral breast carcinomas may be of value in clinical practice and warrant further studies.     

Challenging breast lesions: pitfalls and limitations of fine-needle aspiration and the role of core biopsy in specific lesions

Core biopsy rapidly replaced fine needle aspiration (FNA) over the past decade in evaluation of diseases of the female breast in many centers in the USA. We continue to heavily utilize FNA for the initial evaluation of breast masses in our institution. In this article, we discuss the cytologic and core biopsy findings in challenging breast lesions such as papillary and mucinous proliferations, fibroepithelial neoplasms, and low grade cancers. We specifically focus on the pitfalls and limitations of both diagnostic modalities in these selected specific lesions.     

Expression of Ki-67 and p27(Kip1) in fine-needle aspirates from breast carcinoma and benign breast diseases

Cell atypia in breast fine needle aspiration (FNA) can introduce some diagnostic difficulties. Molecules reflecting proliferative cell potential, such as Ki‐67 and p27^Kip1, can help in recognizing the true biological nature of a cell. Thus, the objective of the study was to analyze the difference in Ki‐67 and p27^Kip1 cell immunoexpression in breast FNA specimens between fibroadenomas, fibrocystic changes (FCC) with atypia, and breast carcinoma. Microscopic analyses of cell cytomorphology and Ki‐67 and p27^Kip1 breast cell immunoexpression were done after standard Pappenheim and immunocytochemical staining (labeled streptavidin‐biotin, LSAB) method in autostainer DakoCytomation TechMate?. The study included 50 patients with breast carcinoma, 20 patients with fibroadenoma, and 20 patients with FCC with atypia. High Ki‐67 and low or absent p27^Kip1 were found in most patients with breast carcinoma, while majority of FCC with atypia were characterized by low Ki‐67 and moderate to high p27^Kip1 cell immunoexpression. Majority of fibroadenomas were associated with low Ki‐67 and low to moderate p27^Kip1 cell immunoexpression indicating progressive decrease in cell cycle inhibition, but still not so high proliferative activity as in carcinoma. However, although statistically significant difference for Ki‐67 and p27^Kip1 was found between breast lesions in our study, the large ranges observed for each marker make them essentially useless for better cytological diagnosis in a single case. Regarding their opposite role in cell cycle, inverse correlation of Ki‐67 and p27^Kip1 was noticed. Poorly differentiated carcinoma cells had mostly high Ki‐67 andlow p27^Kip1 cell immunoexpression. Diagn. Cytopathol. 2011;39:333–340. © 2010 Wiley‐Liss, Inc.     

基底型细胞角蛋白在乳腺导管内增生性病变诊断中的应用

目的比较不同类型基底型细胞角蛋白(CK5、CK34βE12和CK14)在乳腺导管内增生性病变中的辅助诊断价值,并结合肌上皮标记、超微电镜对普通型导管增生的细胞成分进行初步分析。方法参照2003年WHO乳腺疾病分类的诊断标准筛选出28例导管普通型增生(UDH)、10例不典型增生(ADH)和25例导管原位癌(DCIS)。所有病例均进行CK5/6、CK34βE12、CK14、CK8、浕SMA、calponin和p63的免疫组化染色。4例UDH和1例DCIS通过电镜观察其增生细胞的超微结构。结果CK5/6在UDH、ADH和DCIS增生细胞中的阳性表达率分别为92.9%、10.0%和0。CK34βE12和CK14的阳性表达率分别为96.4%和82.1%(UDH)、20.0%和30.0%(ADH)、24.0%和28.0%(DCIS)。所有UDH的增生细胞均不表达浕SMA、calponin和p63。电镜观察显示UDH和DCIS的增生细胞中未发现符合肌上皮超微特征的细胞存在。结论基底型CK有助于UDH和ADH/DCIS的鉴别诊断,其中CK5/6较CK34βE12和CK14特异性更高。免疫组化染色和电镜观察结果支持UDH的增生细胞含有多种成分,包括定向干细胞、腺中间细胞和腺终端细胞等,但未发现具有肌上皮特点的细胞参与其中。     

Morphometric analysis and p63 improve the identification of myoepithelial cells in breast lesion cytology

Nuclear findings, including size, shape, chromatin, or nucleoli, affect the cytological diagnosis and were found to have prognostic significance in several studies based on morphometric analysis. In this study, we investigated whether the nucleus area relates to the appearance of p63-positive myoepithelial cells. The nucleus area was analyzed in 101 breast cancers and 14 benign breast lesions using morphometric analysis and immunohistochemistry for p63. Breast cancer patients were classified into two groups; small nucleus group (n = 19) and large nucleus group (n = 82) based on the average nucleus area of ductal cells. The nucleus area was significantly negative correlating with p63-positive myoepithelial cells (P = 0.031), and there was a significant positive correlation with small cells with hyperchromatic nuclei (P = 0.002). Although small cells with hyperchromatic nuclei were similar to myoepithelial cells, there was no significant correlation with the appearance of small cells with hyperchromatic nuclei and p63-positive myoepithelial cells in breast cancer patients (P = 0.189). Although the identification of normal myoepithelial cells is mainly performed by immunostaining for p63, morphometric analysis of the nucleus area also suggests the existence of p63-positive myoepithelial cells.     

Interobserver variability in the classification of proliferative breast lesions by fine-needle aspiration: Results of the Papanicolaou Society of Cytopathology study

This study evaluates the applicability of the published cytologic criteria in the categorization of proliferative breast lesions by assessing the diagnostic accuracy and interobserver reproducibility of a panel of experts. Twelve breast fine-needle aspiration (FNA) specimens of biopsy-proven nonproliferative breast lesion (NPL) (1 case), proliferative lesions without atypia (PL) (7 cases), proliferative lesion with atypia (PLA) (1 case), and low-nuclear grade ductal carcinoma in situ (DCIS) (3 cases) were selected. Six FNAs were Papanicolaou (PAP) and 6 were Diff-Quik-stained (DQ). Six expert cytopathologists classified the smears using a summary of published criteria as a guideline. All 6 participants rendered the same cytologic diagnosis in 2/12 (16%) cases. The agreement among the 6 raters was low (Kappa = 0.35). Cytohistologic correlation was achieved in 26/72 (36%) FNA diagnoses. The correlation of the PAP-stained cases was better than the DQ: 17/36 (47%) PAP and 9/36 (25%) DQ correlated. Improving the correlation was achieved by amalgamation of NPL and PL into “low risk” and PLA and DCIS into “high risk” categories: 47/72 (65%) FNA diagnoses then correlated with histology [29/36 (81%) PAP and 18/36 (50%) DQ]. We conclude that the cytologic criteria of proliferative breast lesions need to be further defined and assessed. Consideration should be given to minimizing the number of diagnostic categories and adopting a terminology that has a direct effect on patient management. Diagn. Cytopathol. 1998;18:150–165. © 1998 Wiley-Liss, Inc.     

A retrospective study of the diagnostic accuracy of fine-needle aspiration for breast lesions and implications for future use

In recent years, the use of fine-needle aspiration (FNA) in the diagnosis of breast lesions has declined in many institutions. We sought to evaluate the role of FNA for breast lesions and the annual rate of the procedure at our institution over a 4(1/2) year period (May 2002-October 2006). A total of 831 FNAs were performed, with 258 (31%) having histologic follow-up. The number of FNAs obtained was 159 from 5/02 to 4/03, 192 from 5/03 to 4/04, 194 from 5/04 to 4/05, 191 from 5/05 to 4/06, and 95 from 5/06 to 10/06. Each case was placed into one of four categories: nondiagnostic (9%), benign (77.5%), atypical/suspicious (5.5%), or malignant (8%). Surgical tissue was available for 37% of nondiagnostic cases, 22% of benign cases, 80% of atypical/suspicious cases, and 72% of malignant cases. The overall sensitivity and specificity for FNA was 83 and 92% respectively. The overall positive and negative predictive values were 83 and 92% respectively. There were no false-positive cases, indicating a positive predictive value of 100% for a Dx of malignancy. For cases with surgical follow-up, the false-negative rate was 5.4%. Although there is a national trend away from FNAs of breast lesion, this has not been the experience at our institution. Although FNA may not be ideal in the initial evaluation of suspicious lesions, we argue that FNA for clinically benign palpable lesions and recurrent carcinomas has significant value.