Expression of c-myc, erbB-2, p53 and nm23-H1 gene product in benign and malignant breast lesions: coexpression and correlation with clinicopathologic parameters

The aims of this study were to assess the expression of protein products of c-myc, erbB-2, p53 and nm23-H1 gene in benign and malignant breast lesions, to estimate their possible coexpression and to correlate the results of immunohistochemical analysis with various clinicopathologic parameters. The method used was the immunohistochemical detection of the corresponding protein. Expression of c-myc protein was high in both malignant and benign lesions (95% and 100%). Expression of erbB-2 and mutated p53 proteins in malignant lesions was 27% and 34%. These proteins were present in benign lesions as well: 7.8% of benign lesions were positive for erbB-2 protein and 19.6% for p53 protein. The expression of nm23-H1 protein was similar in benign and malignant lesions: 47% and 54%. The coexpression of nm23-H1 and mutated p53 protein was found in 14 carcinomas (16.5%). We found a tendency of negative correlation between the expression of these two proteins. We also found a negative correlation between the size of breast carcinomas and the expression of nm23-H1, a higher proportion of nm23-H1-positive carcinomas in the group of erbB-2-negative, p53-negative carcinomas and a higher proportion of nm23-H1-positive carcinomas in the group of malignant lesions with negative axillary lymph nodes. Our results support the hypothesis that in women with breast cancer the expression of nm23-H1 gene may contribute to more favorable phenotype. We also showed that some changes found in malignant breast tumors such as the presence of mutated p53 protein and the expression of erbB-2 protein may be found in benign lesions as well.     

SIRT6和HIF-1α在三阴性乳腺癌中的表达及临床价值

目的 探索SIRT6基因与缺氧诱导因子（HIF-1α）在三阴性乳腺癌（TNBC）中的表达关系,分析二者相关性及临床价值。方法 收集2017年6月~2018年12月佳木斯大学附属第一医院病理科60例TNBC乳腺癌组织及其对应60例腋窝淋巴结病理组织,并取癌旁正常组织40例。采用免疫组化法检测癌组织、转移淋巴结及癌旁组织中SIRT6、HIF-1α表达,分析二者表达与TNBC乳腺癌临床病理参数、发生发展的关系以及二者表达的相关性。结果 SIRT6在TNBC乳腺癌组织中表达阳性率（80.00%）高于癌旁组织（24.00%）,差异有统计学意义（P＜0.05）,其表达与TNBC乳腺癌组织分化程度,临床TNM分期、转移淋巴结有关（P＜0.05）;在TNBC乳腺癌组织、癌旁组织中HIF-1α的阳性率分别为86.67%、15.00%。且HIF-1α在TNBC乳腺癌组织与癌旁组织表达差异有统计学意义（P＜0.05）。并与TNBC乳腺癌的组织分化程度,临床TNM分期、转移淋巴结相关（P＜0.05）;在TNBC乳腺癌中SIRT6与HIF-1α的表达呈正相关（r=0.823,P＜0.05）。结论 在TNBC乳腺癌中SIRT6与HIF-1α的较高的阳性表达率可能与其发生发展及早期转移有关,检测SIRT6与HIF-1α将有助于了解TNBC乳腺癌的进展及侵袭转移情况,它们有希望为TNBC乳腺癌的早期发现、治疗及预后的评估提供新的研究思路。     

层粘连蛋白及其受体在乳腺癌的表达与转移和预后的关系

目的观察层粘连蛋白（ｌａｍｉｎｉｎ,ＬＮ）和层粘连蛋白受体（ｌａｍｉｎｉｎｒｅｃｅｐｔｏｒ,ＬＮ－Ｒ）与乳腺癌转移和预后的关系。方法用ＬＳＡＢ免疫组化法,检测了１０９例乳腺癌原发灶和３７例淋巴结转移灶组织中ＬＮ和ＬＮ－Ｒ在癌细胞内表达的情况。结果有３２例（２９．４％）原发灶浸润性癌细胞内可见ＬＮ表达,５例（１３．７％）腋窝淋巴结转移灶中癌细胞内有ＬＮ表达。乳腺癌原发灶中有ＬＮ－Ｒ表达者占５５．０４％,淋巴结转移灶中ＬＮ－Ｒ表达占８３．７８％,后者明显高于前者（Ｐ＜０．０５）。６４例随访材料中单独ＬＮ表达者无１例死亡,而有ＬＮ－Ｒ表达者生存期明显短于无ＬＮ－Ｒ表达者（Ｐ＜０．００１）。经临床和病理多因素比例风险回归分析,显示淋巴结转移和ＬＮ－Ｒ是影响患者生存的独立因素,ＬＮ－Ｒ的风险度（４．３７５倍）大于淋巴结转移（２．８１０倍）。结论结果提示ＬＮ－Ｒ表达是导致乳腺癌患者死亡的重要生物学因素之一。     

CD44 expression and axillary lymph node metastasis in infiltrating ductal carcinoma of the breast

Metastasising ability connotes one of the most important life-threatening properties of malignant neoplasms. Recent studies indicate that CD44 proteins, multifunctional cell adhesion molecules which contribute to "homing" of lymphocytes to lymph nodes as well as cell-cell and cell-matrix interactions, are potential markers of tumour progression. However, whether CD44 expression by human tumours contribute to increased metastatic risk remains controversial. In an attempt to clarify its role in breast cancer, we have investigated the correlation between CD44 expression by breast carcinoma and the presence of axillary lymph node metastases. CD44 expression was detected using a standard immunoperoxidase method on formalin-fixed, paraffin-embedded, primary infiltrating ductal breast carcinoma tissues taken from 60 female patients who underwent mastectomy with axillary node clearance. Tumours were graded according to the modified Bloom and Richardson criteria. 62% of patients had histologically-proven lymph node metastasis. 40% of primary cancers exhibited cytoplasmic membrane immunopositivity for CD44. 46% of primary tumours which have metastasied to axillary lymph nodes were CD44 positive whereas 30% of tumours which have not metastasised expressed CD44. CD44 positivity was expressed by 20% of grade 1, 31% grade 2 and 58% grade 3 tumours. Our results suggest that CD44 may have a role in the progression of breast cancer and emphasise the need to investigate its interaction with other mechanisms of cancer advancement.     

吲哚胺2,3-双加氧酶参与乳腺癌患者免疫耐受的研究

目的:研究吲哚胺2,3-双加氧酶(Indoleamine2,3-dioxygenase,IDO)在乳腺癌组织和引流淋巴结中的表达及调节性T细胞在相应组织内的分布,探讨IDO在乳腺癌免疫耐受中的作用机制.方法:收集26例乳腺癌患者的癌组织、癌旁正常乳腺、引流淋巴结和10例乳腺良性病变组织,用RT-PCR法检测IDO mRNA表达,用免疫组织化学法检测IDO和Foxp3蛋白表达.结果:乳腺癌引流淋巴结中IDO mRNA表达水平及IDO表达阳性指数[(19.59±7.65)%]高于原发乳腺癌组织[IDO表达阳性指数(13.16±7.82)%](P<0.05),乳腺癌组织中IDO mRNA表达水平及IDO表达阳性指数高于乳腺良性病变组织[IDO表达阳性指数(3.24±1.30)%]和癌旁正常乳腺组织[IDO阳性细胞指数(2.70±1.53)%](P均<0.05).乳腺癌组织中IDO表达水平与肿瘤临床分期和淋巴结转移相关(P<0.05).乳腺癌引流淋巴结中Foxp3阳性细胞指数[(6.13±2.31)%]高于乳腺原发癌[(3.50±1.04)%],乳腺癌组织中Foxp3阳性细胞指数高于乳腺良性病变[(0.71±0.42)%]和癌旁正常乳腺组织[(0.55±0.34)%](P均<0.05).乳腺癌和引流淋巴结中IDO的表达水平与Treg细胞的分布间均正性相关(r~2=0.449,r~2=0.454,P均<0.05).结论:IDO在乳腺癌细胞中表达增高,并伴随乳腺癌和引流淋巴结中Treg细胞比例升高,提示IDO表达增高有可能通过募集Treg细胞参与肿瘤和引流淋巴结内的免疫耐受.     

Significance of inducible nitric oxide synthase expression in benign and malignant breast epithelium: an immunohistochemical study of 151 cases

The role of calcium independent inducible nitric oxide synthase (iNOS) in breast carcinoma is controversial, and the implications of iNOS expression on prognosis are not known. In this study, we aimed to investigate the significance of immunohistochemical iNOS expression in 100 invasive ductal carcinomas. In addition, 11 normal breast tissues, 20 cases of usual ductal hyperplasias (UDHs) and 20 fibroadenomas were included. We found that 78% of malignant and 75% of benign cases showed iNOS immunoreactivity. However, the intensity and the quantity of iNOS expression were significantly higher in the cancer group when compared with benign breasts (P<0.001), suggesting a role of iNOS in breast carcinogenesis. We were unable to show a correlation between iNOS expression and tumor grade, axillary lymph node status, and estrogen receptor expression. In 50 axilla negative cases having 5–12 years follow-up, disease free survival (DFS) rate was significantly lower in cases showing strong iNOS expression (P=0.05). As strong iNOS expression was correlated with short DFS, we concluded that further studies would be necessary to elucidate if iNOS expression might be a useful prognostic marker in breast carcinoma, especially in the axilla negative group.Part of this study was presented at XXIVth International Congress of the International Academy of Pathology, Amsterdam, The Netherlands on 5–10 October 2002 as a poster presentation.     

Benign axillary epithelial lymph node inclusions—a histological pitfal

We present two cases of axillary epithelial lymph node inclusions in the presence of benign and malignant breast disease. Although the presence of lymph node inclusions is well recognized at other sites in the body, their presence in the axillary nodes of women with breast disease necessitates close attention. This is particularly true in women with malignant breast disease as misinterpretation may lead to inappropriate treatment.     

Immunohistochemical staining of Ha-ras oncogene product in normal, benign, and malignant human pancreatic tissues

We examined immunohistochemical staining with Ha-ras oncogene product in normal, benign, and malignant human pancreatic tissues. In cases of pancreatic cancer, its relation to histologic type was evaluated. Serous cystadenoma and atypical acinar cell nodules did not react with the Ha-ras oncogene product, and ductal cells and acinar cells in normal pancreas showed lower immunoreactivity than other benign or malignant lesions. However, the positive rate of islet cells in normal pancreas was almost the same in cases of pancreatic cancer. Strongest positivity was observed in cases of chronic pancreatitis, and islet cell tumors also showed high positive staining rates. In pancreatic cancer, the positive rate of well-differentiated adenocarcinomas was rather higher than that of poorly differentiated adenocarcinomas. Our study indicates that the Ha-ras oncogene p21 product does not correlate with neoplastic transformation in human pancreas, is not a useful marker for differentiating benign and malignant lesions, and cannot be used to determine the origin of differentiation in the pancreas.     

Dickkopf-1 (Dkk1) protein expression in breast cancer with special reference to bone metastases

Dysregulation of the Wnt inhibitor dickkopf-1 protein (Dkk1) has been reported in a variety of cancers. In addition, it has been linked to the progression of malignant bone disease by impairing osteoblast activity. This study investigated serum- and tissue levels of Dkk1 in breast cancer patients with- or without bone metastases. Serum Dkk1 levels were measured by ELISA in 89 breast cancer patients and 86 healthy women. Tissue levels of Dkk1 and β-catenin, a major downstream component of Wnt transduction pathway, were tested with immunohistochemical staining in 143 different tissues, including adjacent non-tumoral breast tissues, primary breast tumours, lymph nodes metastases, and bone metastases. Serum levels of Dkk1 were significantly increased in breast cancer patients without metastases compared with healthy controls and even more increased in patients with bone metastases. Tissue expression of Dkk1 was positive in 70% of tested primary breast cancer tissues and demonstrated significant correlation with histological type and PR status. Less frequent expression of Dkk1 was found in lymph nodes metastases and bone metastases compared with adjacent non-tumoral breast tissues and primary breast tumours. Tissue expression of β-catenin was positive in the vast majority of all tested tissue types indicating activated Wnt/β-catenin signalling. Our results suggested that Wnt/β-catenin signalling in breast tumours and their secondary lymph nodes- and bone metastases is dysregulated and this could be related to aberrant Dkk1 expression levels. Hence, Dkk1 protein might provide insights into the continued development of novel comprehensive and therapeutic strategies for breast cancer and its bone metastases.     

埃兹蛋白在乳腺癌中的表达及其临床意义

目的 通过检测埃兹蛋白在浸润性乳腺癌、乳腺导管内癌及正常乳腺组织中的表达,探究埃兹蛋白在乳腺癌组织中的表达的临床意义。方法 随机选取南昌大学第一附属医院40例浸润性乳腺癌患者及32例乳腺导管内癌患者,另外选择同期来我院健康体检者22例。通过免疫组化链霉亲和素-过氧化物酶法（S-P）法分别检测埃兹蛋白在22例正常乳腺组织、32例乳腺导管内癌组织和40例浸润性乳腺癌组织中的表达,参照病理结果进行分析。结果 埃兹蛋白在40例浸润性乳腺癌的阳性表达率为72.50%、在32例乳腺导管内癌的阳性表达率为40.63%、在正常乳腺组织中的阳性表达率13.64%,浸润性乳腺癌中埃兹蛋白的表达高于其他两种组织,差异具有统计学意义（P＜0.05）;埃兹蛋白在G3期乳腺癌组织中的表达高于在G1~G2期乳腺癌组织中的表达,差异具有统计学意义（P＜0.05）;埃兹蛋白在淋巴结转移患者中的阳性表达率高于在无淋巴结转移的患者中的阳性表达率,差异统计学意义显著（P＜0.01）;埃兹蛋白在＜40岁的患者中的阳性表达率低于在年龄≥40岁的患者中的表达率,差异统计学意义显著（P＜0.01）;埃兹蛋白在PR阳性患者中的阳性表达率低于在PR阴性患者中的表达率,差异具有统计学意义（P＜0.05）;乳腺癌患者中埃兹蛋白的表达与组织学分级、腋窝淋巴结转移及年龄呈正相关（P＜0.05）,与PR呈负相关（P＜0.05）;埃兹蛋白在浸润性润腺癌中的表达与乳腺癌临床分期、脉管侵犯、肿瘤大小、患者月经状态以及Her-2、ER、Ki-67的表达无相关性（P＞0.05）。结论 埃兹蛋白在乳腺癌和正常乳腺组织中的表达不同,对判断乳腺肿瘤性质具有重要参考意义;此外,埃兹蛋白能够帮助判断乳腺癌的转移潜能和预后,可能启发乳腺癌新的靶向治疗。     

mRNA markers of breast cancer nodal metastases: comparison between mammaglobin and carcinoembryonic antigen in 248 patients

Histological detection of axillary lymph node metastases is still the most valuable prognostic parameter for breast cancer, but about 30% of node-negative patients relapse within five years, suggesting that current methods are inadequate for identifying metastatic disease. More sensitive, PCR-based methods for the detection of metastatic cells are now available, enabling the amplification of cancer cell-specific mRNA messages by the RT-PCR assay. An ideal tumour marker, consistently expressed in tumour samples and not at all in normal lymph nodes, remains to be identified. The present study first investigated the expression of seven mRNA markers, CEA, CK19, c-Met, mammaglobin, MUC-1, beta1-->GalNAc-T and p97, selected on the basis of their previously reported specificity for breast cancer cells. Eighteen lymph nodes were examined from patients without tumours. Only mammaglobin mRNA and CEA mRNA were not expressed in normal nodes. All of the other markers showed a band of expression in 17%-55% of cases, indicating that they are not breast cancer-specific. CEA mRNA and mammaglobin mRNA expression could be detected in 15/20 (75%) and 19/20 (95%) primary breast carcinomas, respectively. The expression of mammaglobin mRNA and CEA mRNA was then compared in axillary lymph nodes from 248 consecutive breast cancer patients, 89 with histologically documented lymph node metastasis and 159 without histological evidence of metastatic disease. Ninety-seven per cent of the patients with histologically involved nodes showed expression of mammaglobin mRNA, whereas CEA mRNA was expressed in 79% of these cases. In the group of patients with histologically negative lymph nodes, 46 (29%) and 32 (20%) were found to be positive for mammaglobin and CEA expression, respectively, indicating the presence of metastases not detected by routine histological examination of one lymph node section. These results show that both mammaglobin RT-PCR and CEA RT-PCR are useful tools for the detection of breast cancer metastases in axillary lymph nodes. The detection sensitivity of the mammaglobin RT-PCR is far superior to that of the CEA RT-PCR, allowing the diagnosis of occult metastases in nearly one-third of cases.     

High expression of REGγ is associated with metastasis and poor prognosis of patients with breast cancer

REGgamma (REGγ) has been recently found in several types of human cancer, however, its clinical significance in metastasis and prognosis of breast cancer remains unknown. In this study, immunohistochemical staining and western blot analysis were performed to evaluate REGγ expression in both mouse and human breast cancer specimens. We found that in MMTV-PyMT mice, 14 out of 20 (70%) mouse mammary carcinomas were REGγ positive, which was significantly higher than control (0/20, 0%, P < 0.001) and lower than metastatic lung tumour (20/20, 100%, P = 0.027). Further investigation for REGγ expression in 136 human breast cancer tissues with the paired peritumoural normal breast tissues and 140 breast benign disease tissue samples showed that REGγ was undetectable in normal breast tissues and nonmetastatic axillary lymph nodes (ALNs), whereas 111 out of 136 (81.6%) breast cancer tissue samples were REGγ positive, which was significantly higher than breast benign disease tissues (9/140, 6.4%, P < 0.001) and lower than metastatic ALNs (116/116, 100%, P < 0.001). The 5-year disease-free and overall survivals of patients with negative/low level of REGγ were significantly higher than those of patients with high level of REGγ (P < 0.05). Cox regression analyses further indicated that REGγ could serve as a novel independent prognostic factor for breast cancer (OR = 4.369, P = 0.008). Our results suggest that the high expression of REGγ might predict metastasis and poor prognosis in breast cancer.     

定量RT—PCR法检测乳腺癌前哨淋巴结CK19的研究

目的探讨定量RT—PCR法检测细胞角蛋白（CK19）在乳腺癌前哨淋巴结（SLN）中的表达，提高前哨淋巴结活检中微转移的检出率。方法采用常规病理检查法（HE染色）和定量RT—PCR法检测了40例乳腺癌患者SLN的CK19的表达量，同时选取10例来源于胃肠道的良性病变淋巴结作为定量RT—PCR检测的对照组。结果CK19在良性病变的淋巴结中没有表达。常规病理检查的敏感度为42．9％（9／21），假阴性为57．1％（12／21）．准确率为70．0％（28／40）。定量RT—PCR法检测出常规病理未检出的微小转移病例12例，敏感度为95．2％（20／21），假阴性为4．8％（1／21），准确率为97．5％（39／40）。结论前哨淋巴结活检可有效判断乳腺癌腋淋巴结转移状态，应用定量RT—PCR法检测CK19在SLN中的表达，可提高敏感度及准确率。     

人乳腺癌组织中CD1C+树突状细胞免疫组化研究

目的观察树突状细胞在人乳腺癌组织中的形态学变化。方法利用CD1C+作为树突状细胞(dendriticcell,DC)的特征性标志分子,用免疫组织化学方法观察20例人乳腺癌组织及癌旁相对正常组织中树突状细胞的分布和形态学变化,同时检测p53蛋白的表达。结果乳癌组织中树突状细胞CD1C+阳性面数密度(4.55±1.32)和平均光密度值(0.13±0.03)明显低正常乳腺组织面数密度(11.16±5.32)和平均光密度值(0.27±0.07,P<0.05);乳癌组织DC数量较正常乳腺组织明显减少(P<0.05)。20例乳腺癌中,p53蛋白阳性表达率为60%;正常组织为阴性。结论树突状细胞及其表达的p53蛋白可能参与肿瘤免疫调节作用。     

Primary small cell (oat cell) carcinoma of the breast: report of a case and review of the literature

A case of primary small cell (oat cell) carcinoma of the breast in a 41-year-old woman is presented. The patient was alive and well without disease 16 months after modified radical mastectomy and subsequent chemotherapy. The tumor cells revealed morphologic similarity to oat cell carcinoma of the lung and immunohistochemical expression of neuroendocrine markers. In ultrastructural examination, the tumor cells had neurosecretory granules. Review of nine previously reported cases and this case of primary small cell carcinoma of the breast has revealed that this type of tumor shows prominent vascular invasion, frequent lymph node metastasis, infrequent expression of estrogen receptor, and also very poor prognosis. Immunohistochemical study for the c-kit proto-oncogene product, which has been reported to be a specific marker for pulmonary small cell carcinoma, demonstrated positive reactivity in approximately 80% of the tumor cells of this case, which is the first report according to our knowledge. The expression of c-kit might be some aid to the diagnosis of primary small cell carcinoma of the breast.     

CD44(+)/CD24(-) cells are transit progenitors and do not determine the molecular subtypes and clinical parameters in breast carcinomas

CD44(+)/CD24(-) cells have been associated with breast cancer stem/progenitor cell features. However, the status of this phenotype cells in normal, benign and malignant breast tissues has not been studied, and the clinical correlation of this subpopulation in breast cancer is not fully understood. The present study sought to identify these cells in a series of normal, benign, and malignant breast tissues and explore their correlation to the molecular subtypes of breast carcinoma and conventional pathological features. Double-staining immunohistochemistry (DIHC) of CD44 and CD24 was performed on 30 normal breast tissues, 30 breast fibroadenomas (FA), 60 breast invasive ductal carcinomas (IDC), and 3 breast cancer cell lines (MCF-7, MDA-MB-468, and MDA-MB-231). In the normal breast tissues and FAs, three phenotypes were observed including CD44(+)/CD24(+), CD44(+)/CD24(-), and CD44(-)/CD24(-) cells. In the IDCs, CD44(-)/CD24(+) cells were detected, in addition to the three aforementioned phenotypes. The strong positive rate (+++, incidence >60%) of CD44(+)/CD24(-) was significantly increased from normal breast tissue, FAs to IDCs (0.0%-->6.7%-->21.7%). However, the CD44(+)/CD24(-) cells didn't correlate with ages of patients, lymph node metastasis, tumor size, molecular subtypes, and the expression of ER, PR, HER-2, PS2, Bcl-2, nm23. The proportion of CD44(+)/CD24(-) cells in MCF-7, MDA-MB-468, and MDA-MB-231 was about 1, 5, and 80%, respectively. The results indicate that the CD44(+)/CD24(-) cells are transit progenitors and have no association with the molecular subtypes and clinicopathological parameters in the IDCs.