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。     

Myoepithelial Cells in Lobular Carcinoma in Situ of the Breast: A Parallel Immunocytochemical and Ultrastructural Study

The distribution and morphology of myoepithelial cells in one case of normal breast and in 4 cases of lobular carcinoma in situ (LCIS) were studied by immunoperoxidase and immunofluorescence with antiactin antisera and by electron microscopy. The actinrich cells appeared to lay flattened on the basement membrane, or perpendicular to it, or appeared dislodged toward the center of the neoplastic ductules in the different cases.

It is proposed that the presence of actin-rich cells inside the neoplastic aggregates represents a process of colonization of lobular carcinoma by myoepithelial cells dislodged from the basement membrane; a differentiation of some neoplastic cells toward myoepithelial elements might also be taken into consideration. The features here described could identify a preinvasive stage of LCIS.     

Ultrastructural Localization Of Endogenous Peroxidase In The Human Thyroid Gland Under Normal And Hyperfunctioning Conditions

Ultrastructural localization of human thyroid peroxidase in normal and hyperfunctioning states has been studied. Peroxidase activity was visualized ultrastructurally with a cytochemical reaction using 0.05% diaminobenzidine-tetrahydrochloride, and hydrogen peroxide with a final concentration of 0. 0025%, 0.005%, or 0.01%, respectively, and 2% osmium tetroxide. Reaction product demonstrating the enzyme was clearly observed in cell organelles with a final concentration of hydrogen peroxide at 0.0025% or 0.005%. In the normal portion of thyroid, follicular cells demonstrated the reaction product, located mainly in rough-surfaced endoplasmic reticulum and perinuclear cisternae and in addition a small amount of the product was found in Golgi cisternae and small vesicles dispersed throughout the subapical cytoplasm. The reaction product significantly increased in amount in the cell organelles described above and in addition, a characteristic heavy deposition of the product was observed at the external surface of the microvilli in the follicular cells of thyroid obtained from the patients with treated Basedow's disease. ACTA PATHOL. JPN. 34: 553–561, 1984.     

Cardiac Neuroendocrine Tumor with Absence of Sustentacular Cells: Immunohistochemical and Ultrastructural Findings

A 68-year-old male presented with increased shortness of breath and intermittent chest pain. Cardiac catheterization along with echocardiogram imaging demonstrated 3-vessel coronary artery disease with severe left ventricular dysfunction and critical aortic stenosis. During coronary artery bypass surgery, a tumor was identified at the sulcus between the aorta and the right atrial appendage. This highly vascular tumor extended over the right coronary artery, prompting biopsy and further resection. Light microscopic evaluation showed packets of uniform round neoplastic cells without evidence of necrosis or increased mitotic figures. Immunohistochemical studies were positive for synaptophysin and weakly positive for chromogranin. Pankeratin, S-100, and GFAP stains were all negative. Ultrastructural examination of the neoplasm demonstrated numerous electron-dense secretory granules within the cytoplasm of the tumor cells. These secretory granules varied in size from 60 to 210?nm, with halos encircling many of them, suggesting the likelihood of epinephrine or norepinephrine granules. Within the specimen, abundant vascular spaces were identified, but no sustentactular cells were present. The patient agreed to undergo genetic testing, and a mutation in the succinate dehydrogenase complex subunit B (SDHB) was identified. This confirmed the molecular diagnosis of hereditary paraganglioma/pheochromocytoma syndrome. Now two years out from surgery, this patient continues to be asymptomatic in spite of the fact that his tumor was only partially resected.     

Myoepithelial Carcinoma Arising in a Benign Myoepithelioma: Immunohistochemical, Ultrastructural, and Flow-Cytometrical Study

A case of myoepithelial carcinoma arising in a benign myoepithelioma of the minor salivary gland in a 71-year-old patient is reported. The tumor presented initially on the palate and had been diagnosed as “benign lesion” 40 years before. It recurred 22, 36, and 40 years after initial presentation, and a similar histopathological diagnosis was rendered. One year after the last recurrence, the tumor recurred showing typical changes of malignant transformation, and the diagnosis was malignant myoepithelioma. The light microscopy and ultra-structural features of the initial tumor were typical of plasmocytoid myoepithelioma. There were abundant round cells and rare spindle cells with uniform dispersed filaments, sometimes arranged in parallel streams without evidence of dense bodies. These cells showed micropinocytotic vesicles along the cell membrane with poorly developed intercellular junctions and were surrounded by a basal membrane. The malignant counterpart showed fewer plasmocytoid cells and a rather epithelial pattern with marked nuclear pleomorphism and formation of small, or rarely large, glandular lumina. The immunohistochemical features were similar for the benign and malignant tumors, with positivity for S-100 protein, vi-mentin, cytokeratins, and CAM 5.2, and were negative for GFAP, muscle-specific actin, CEA, and desmin. Flow cytometry showed a change in the DNA content profile. The benign myoepithelioma had a diploid DNA content with a low S-phase fraction of 3.9% and proliferative index of 9.1%, while the myoepithelial carcinoma had an evident aneuploid DNA stem line and an increased S-phase fraction of 8.3% with a proliferative index of 18.1%.     

抗人乳腺癌单克隆抗体HD1的特性

HD1是由1株抗人乳腺癌杂交瘤腹水扩增,亲和层析纯化得到的单克隆抗体,分泌抗体(IgG)的蛋白含量:上清为10μg/ml,腹水为1.5mg/ml。用ABC法检测,HD1与3株人乳腺癌细胞系呈阳性反应,而与胃癌、结肠癌等细胞和正常成纤维细胞、淋巴细胞呈阴性反应。组织化学染色显示对部分乳腺癌石蜡切片为阳性,而对多种正常组织为阴性。HD1对3株乳腺癌细胞系的亲和力在1.3×10~(-8)～2.7×10~(-8)mol/L之间。免疫印迹实验表明,HD1可使乳腺癌细胞MCF-7和475-A细胞膜Mr为60000的抗原组分染色,使另一株乳腺癌细胞BCaP-37 Mr为56000和94000的组分着色。对3株乳腺癌细胞有抗体介导的细胞毒效应(18％～27％)。以上的研究有助于对HD1性质及其相对应抗原的了解,为抗乳腺癌单抗的应用提供了可靠的依据     

Phosphosulindac (OXT-328) Selectively Targets Breast Cancer Stem Cells In Vitro and in Human Breast Cancer Xenografts

Pharmacological targeting of breast cancer stem cells (CSCs) is highly promising for the treatment of breast cancer, as the small population of CSCs appears responsible for tumor initiation and progression and also for resistance to conventional treatment. Here we report that the novel phosphosulindac (OXT-328, PS) selectively and effectively eliminates breast CSCs both in vitro and in vivo. PS reduced cell proliferation and induced apoptosis in various breast CSCs. Breast CSCs are resistant to conventional cancer drugs but are sensitive to PS. Long-term treatment of mixtures of cultured breast CSCs and breast cancer cells with PS preferentially eliminated the CSCs. PS impaired the ability of CSCs to form mammospheres and markedly suppressed the expression of CSC-related genes. More importantly, PS prevented by half (p =.06) the formation of tumors initiated by CSCs in immunodeficient mice, and inhibited by 83% (p <.05) the growth of already formed breast cancer xenografts, reducing the proportion of CSCs in them. PS suppressed the Wnt/β-catenin pathway by stimulating the degradation of β-catenin and its relocalization to the cell membrane and also blocked the epithelial-mesenchymal transition and the generation of breast CSCs. These results indicate that PS has a strong inhibitory effect against breast cancer, acting, at least in part, by targeting CSCs through a signaling mechanism involving Wnt signaling. STEM Cells2012;30:2065-2075.     

Malignant Myoepithelioma (Myoepithelial Carcinoma) of the Breast: An Ultrastructural and Immunocytochemical Study

This report describes the light (LM) and electron microscopic (EM) features and the results of an indirect immunofluorescence study (IF), the latter using monoclonal and monospecific antibodies to cytoskeletal proteins, of a malignant, invasive and metastatic breast myoepithelioma. A 53-year-old female underwent mastectomy for a large necrotic mammary tumor that had invaded the overlying skin. By LM, the neoplasm was composed of interlacing bundles of large, elongated and interspersed stellate cells with acidophilic cytoplasm. The neoplastic cells displayed a moderate degree of anaplasia, high mitotic activity, and strong tendency for necrosis. Stromal desmoplasia was marked, especially toward the center of the neoplasm. By IF, the tumor cells revealed bright cytoplasmic fluorescence with antibodies to actin, prekeratin, and cytokeratin. A few scattered spindle cells, which stained with the anti-vimentin and anti-actin antibodies, most likely represented stromal myofibroblasts. The anti-desmin reaction was negative. By EM, the neoplasm was composed of variably differentiated, elongated and stellate myoepithelial cells connected by desmosomes, enveloped by remnants of basal lamina, and containing pinocytotic vesicles, a well-developed rough endoplasmic reticulum, large Golgi areas, aggregates of intermediate filaments that were often arranged in dense curvilinear bundles (tonofilaments), and bundles of microfilaments with fusiform, dense bodies. The combined LM, EM, and IF study of this mammary tumor establishes its myoepithelial origin and, thus, identifies it as myoepithelial carcinoma distinct from other spindle cell breast tumors. This neoplasm was locally invasive and cytologically malignant; moreover, its malignancy was further confirmed by the development of lung and pleural metastases.     

重组IL-12对人乳腺癌细胞自噬的研究

目的 探讨重组人IL-12（rIL-12）对乳腺癌细胞自噬的影响。方法 两株乳腺癌细胞（MDA-MB-231和MCF7）分别用rIL-12处理,经免疫蛋白印迹技术和细胞免疫荧光技术检测其自噬微管相关蛋白轻链3（LC3）的变化,以观察自噬情况;另外,用透射电镜观察rIL-12处理乳腺癌细胞前后自噬小体的变化。分别用胰岛素样生长因子1（IGF-1）激活磷脂酰肌醇-3激酶/蛋白激酶B（PI3K/Akt）通路,再加入rIL-12处理后,蛋白免疫印迹法检测相关通路蛋白PI3K/Akt, 哺乳动物雷帕霉素靶点（TOR）磷酸化变化及LC3的表达。结果 与空白组相比,rIL-12组细胞的自噬标志蛋白LC3表达增高,差异有统计学意义（P＜0.05）,且增加程度呈时间和浓度依赖性;荧光显微镜观察自噬体形成的结果显示,与空白组相比,rIL-12处理组的细胞核周点状聚集的绿色荧光增多;使用电镜观察到rIL-12处理组明显有自噬小体形成。与rIL-12组相比,IGF-1+rIL-12组的LC3Ⅱ表达减少,差异有统计学意义（P＜0.05）。结论 rIL-12可以激活乳腺癌细胞自噬,并通过抑制PI3K/Akt信号通路的机制,促进自噬标志蛋白LC3,特别是LC3Ⅱ的表达。     

人乳腺癌组织淋巴管的超微结构

目的 观察人乳腺癌组织毛细淋巴管的超微结构特点,探讨乳腺癌细胞淋巴道转移机理.方法 取20例人乳腺癌术后标本,按癌中心区、周边区及正常区切取组织,半薄切片,光镜定位,透射电镜观察乳腺癌组织毛细淋巴管超微结构.结果 乳腺癌组织中心区未见淋巴管,癌周边区和正常区组织内存在毛细淋巴管.与正常区比较,周边区组织内淋巴管数量增多,管腔扩大,形态不规则;淋巴管内皮细胞连接开放增多,并可见部分内皮细胞破裂溶解,管壁不完整;毛细淋巴管内皮细胞变性,高尔基体变形,粗面内质网脱颗粒,溶酶体数量增多和形态改变.结论 乳腺癌周边区组织内毛细淋巴管内皮细胞的超微结构有改变,提示癌细胞对内皮细胞有破坏、溶解作用;癌细胞可能以通过癌周边区毛细淋巴管内皮细胞连接的开放和毛细淋巴管内皮破坏两种方式侵袭淋巴管而发生淋巴道转移.     

单个乳腺癌细胞表面增强拉曼光谱研究

将内吞进入细胞内部的金纳米颗粒作为表面增强拉曼散射光谱的活性增强基底,通过共聚焦显微拉曼光谱仪测试单个人乳腺癌细胞(MCF-7)的常规拉曼光谱和表面增强拉曼光谱,并进行了初步峰位归属分析及不同拉曼峰强度柱状图对比分析。实验结果表明,内吞金纳米颗粒的单个MCF-7细胞的表面增强拉曼光谱相比于常规拉曼光谱包含更多的细胞生化物质拉曼信号,并且谱线强度有显著增强。由于细胞中各种生物分子在结构和含量上的变化可以反映细胞代谢及病理变化,因此,以具有高灵敏度的金纳米颗粒作为生物分子探针可能成为探测细胞内部组分信息的一种有效的方法,基于内吞金纳米颗粒的单个MCF-7细胞SERS技术在乳腺癌的诊断和治疗方面具有广阔的应用前景。     

Intraductal Papilloma of the Male Breast: An Ultrastructural and Immunohistochemical Study

A case of intraductal papilloma of the male breast was studied by electron microscopy and immunohistochemistry. The major components of this lesion were the epithelial and myoepithelial cells. Intermediate cells showing ultrastructural features of both cell types were also observed. Squamous metaplasia was noted in many areas. Numerous intranuclear helioid inclusions were seen in the tumor cells. The features of this lesion are similar to those of papillomas of the female breast.     

Intraductal Papilloma of the Male Breast: An Ultrastructural and Immunohistochemical Study

A case of intraductal papilloma of the male breast was studied by electron microscopy and immunohistochemistry. The major components of this lesion were the epithelial and myoepithelial cells. Intermediate cells showing ultrastructural features of both cell types were also observed. Squamous metaplasia was noted in many areas. Numerous intranuclear helioid inclusions were seen in the tumor cells. The features of this lesion are similar to those of papillomas of the female breast.     

Mucinous Colorectal Adenocarcinoma with Signet-Ring Cells: Immunohistochemical and Ultrastructural Study

A primary mucinous colorectal adenocarcinoma tissue with signet-ring cells, as revealed after histological evaluation, was examined ultrastructurally. The authors also analyzed the immunohistochemical data of the tissue for serotonin, vasoactive intestinal polypeptide (VIP), bombesin, somatostatin, and glucagon, using the peroxidase anti-peroxidase (PAP) method and the immunogold labeling method for light and electron microscope, respectively. Electron microscopically mucinous adenocarcinoma was characterized by the formation of small lumen. Adenocarcinoma cells were full of mucous granules of varying electron density, providing a good environment for the tumor cells to grow. They also exhibited a significant loss of microvilli and intracytoplasmic junctions, which could allow the cells to disseminate. Signet-ring cells were located in the basal site of the ducts or in the lamina propria and appeared neoplastic, with mucin accumulation intracellularly and an eccentric crescent-shaped nucleus. The cytoplasmic organelles were decreased and at the periphery of the cell. The PAP method demonstrated that these cells were strongly positive for bombesin and also positive for vasointestinal polypeptide (VIP). The immunogold method detected bombesin immunoreactivity in the vacuoles as well as in other cytoplasmic membranes, whereas VIP was localized mainly in the plasma membrane. The location of signet-ring cells combined with the immunoreactivity for bombesin and VIP indicated that signet-ring cells were of neuroendocrine origin and probably dedifferentiated enterochromaffin-like endocrine cells. These findings have implications for understanding the biological behavior of these composite malignant tumors and could help in the knowledge of the origin of signet-ring cells.     

Apoptosis Induction of Centella Asiatica on Human Breast Cancer Cells

The present study evaluated the ability of methanolic extract of Centella asiatica (Linn) Urban (Umbelliferae) to induce apoptosis in different cancer cell lines. MCF-7 cells emerged as the most sensitive cell line for in vitro growth inhibitory activity. C. asiatica extract induced apoptosis in MCF-7 cells as indicated by nuclear condensation, increased annexin staining, loss of mitochondrial membrane potential and induction of DNA breaks identified by TUNEL reactivity. It is possible that the use of C. asiatica extract as a component in herbal medicines could be justifiable.     

Induction Of Megamitochondria In The Rat Liver By N-Propyl Alcohol And N-Butyl Alcohol

Propyl alcohol and butyl alcohol had similar effects to ethyl alcohol on ultrastructure of liver mitochondria. Rats were given 32% ethyl alcohol, 32% n-propyl alcohol, and 6.9% n-butyl alcohol in drinking water for up to three months. After one month, mitochondria in hepatocytes obtained from the experimental animals became elongated, constricted or cup-shaped with scanty cristae. After two months, mitochondria in some hepatocytes became gigantic. In extreme cases, the megamitochondria exceeded 10 μm in diameter. Coupling efficiencies of hepatic mitochondria obtained from alcohol-fed animals were well preserved despite their drastic morphologic changes. ACTA PATHOL. JPN. 34: 471–480, 1984.     

Immunohistochemical Demonstration of Cell Proliferation and Estrogen Receptor Status in Human Breast Cancer

Cell proliferation and estrogen receptor (ER) status was investigated in 45 invasive ductal carcinomas of the breast by immunohistochemical methods using monoclonal antibodies Ki 67 (anti-human proliferating cell antibody) and ER ICA. The results were assessed on the basis of nuclear staining intensity and the percentage of positively stained tumor cell nuclei (index score). There was a significant inverse correlation between the Ki 67 and ERICA index scores, although 4 cases showed high index scores for both markers. We conclude that ER positive cells do not always have low proliferation activity, which may be one of the reasons why endocrine therapy is not effective against all ER positive breast cancers. Acta Pathol Jpn 40: 902 907, 1990.     

Effect of Norcantharidin on the Human Breast Cancer Bcap-37 Cells

Norcantharidin (NCTD), a chemically modified form of cantharidin, is a potential anticancer drug. In this study, the effects of NCTD on the cellular viability, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and DNA damage in the human breast cancer cell line Bcap-37 were investigated with confocal and fluorescence microscopy. The cell cycle was further analyzed using the CellQuest software of a Becton-Dickinson FACS flow cytometer. The results indicated that the cellular viability was decreased with the growing concentrations of NCTD and time exposure. Moreover, the fluorescence intensity of ROS was increased, whereas the MMP was decreased in Bcap-37 cells with the growing concentrations of NCTD. NCTD induced a dose-dependent DNA damage and reduced the G1 peak in Bcap-37 cells. The G2/M peak of Bcap-37 was also decreased by the higher concentration of NCTD.     

Membrane Fusion In Mitochondria Ii. An Ultrastructural Interpretation Of Cristae-Enriched Megamitochondria Induced By Cuprizone

Ultrastructure of megamitochondria in mouse hepatocytes induced by cuprizone was analyzed in the light of the fusion of inner mitochondrial membranes. Hepatic parenchymal cells of mice fed with a 0.5% cuprizone-diet for 7–8 days were characterized by megamitochondria with numerous cristae including disoriented, shorter ones than those of the control and those in vesicular configuration. After feeding the animals with cuprizone for 15–16 days, megamitochondria increased their sizes further and the number of cristae decreased drastically. Numbers of cristae based on a unit-mitochondrial area were 41.3$pL10.8 for the former type of cristae-enriched megamitochondria and 3.8 $pL 1.8 for the latter type of megamitochondria with the value of 23.9 $pL 6.2 for the control. Considering the content of cytochromes in megamitochondria previously reported, these results might suggest that the fusion and defusion of inner mitochondrial membrane take place along with the fusion of outer membranes during the megamitochondrial formation. ACTA PATHOL. JPN. 34: 481–488, 1984.