乳腺肿瘤基膜与间质圆形细胞和S_（－100）阳性细胞浸润关系的研究

应用免疫组化方法以抗层粘连蛋白（ＬＮ），Ⅳ型胶原（ＣｏｌｌⅣ）抗体分别对９８例各类乳腺癌和乳腺良性病变进行了研究。发现ＬＮ和ＣｏｌｌⅣ分布在瘤细胞与间质接邻处，两者的染色结果基本一致；在有密集圆形细胞浸润的癌巢周围，尤见于导管内癌，可见基膜染色变浅或中断；１９／６２例（３０．７％）间质树突状细胞阳性和２／３１例（６．５％）间质树突状细胞阴性连续线状基膜阳性（Ｐ＜０．０５）。本文认为间质中的炎症细胞浸润和树突状细胞分布与基膜改变有密切关系，但尚需深入研究。     

食管鳞癌层粘连蛋白表达的意义

应用兔抗鼠层粘连蛋白抗体、ＰＡＰ免疫组化方法研究８０例食管鳞癌层粘连蛋白的表达结果显示：癌旁正常上皮、单纯性增生及不典型增生上皮层粘连蛋白均呈连续线状，原位癌呈线状，但可见局部中断，浸润癌层粘连蛋白表达呈多种形式；分化好的团块型及网状型在癌巢周围呈线状，但呈明显不规则和不连续，分化差的细索型癌则常呈碎片状，在癌浸润前缘有时见不定形阳性物质堆积。分析层粘连蛋白表达癌的分级，血管、神经侵犯及淋巴结转移     

纤维粘连蛋白在乳腺癌中的表达及与淋巴结转移的关系

目的 研究纤维粘连蛋白（ＦＮ）在乳腺浸润性癌中的表达及其与同侧腋窝淋巴结转移的关系。方法 应用免疫组化（ＳＰ）法检测５０例浸润性癌底膜及间质ＦＮ的表达。结果 癌巢周围基底膜ＦＮ的表达与癌细胞分化程度及淋巴结转移有关,而间质ＦＮ的表达与以上二者均无关,结论癌巢周围基底膜ＦＮ的表达对乳腺癌预后的评估有意义。     

胰腺癌细胞凋亡相关基因的表达及相互关系

本文采用免疫组化ＳＰ法检测４８例胰腺癌和１５例胰腺癌转移灶石蜡标本中ｂｃｌ－２、ｐ５３、ｃ－ｍｙｃ的表达。结果显示：ｂｃｌ－２蛋白在胰腺癌组明显高于胰癌转移灶组；无转移的、高分化的胰腺癌明显高于有转移的和低分化胰腺癌。Ｐ５３蛋白在胰腺癌组有明显高于胰腺良性病变组，胰腺高分化癌明显低于分化癌。ｂｃｌ－２和Ｐ５３蛋白在胰腺癌组中呈负相关。     

P21和P53蛋白在食管癌表达的免疫组化研究

作者用免疫组化ＡＢＰＡＰ法观察４０例食管癌及癌旁组织的ｒａｓ癌基因产物Ｐ２１和癌基因Ｐ５３蛋白的表达。结果显示：Ｐ２１和Ｐ５３蛋白阳性表达分别有２４例和８例（阳性率为６０％和２０％）、。高分化鳞癌以Ｐ２１蛋白表达为主，低分化鳞癌则以Ｐ５３蛋白表达较多；而粘液表皮样癌和腺鳞癌几乎均有超表达现象。癌旁粘膜两种蛋白表达的阳性率明显低于癌组织。Ｐ２１和Ｐ５３蛋白阳性病例，其淋巴结转移率有显著的差异（Ｐ＜：０．０５）。作者认为，在食管癌病人中，检测其癌组织的Ｐ５３蛋白比Ｐ２１蛋白更加有助于了解其生物学特性及预测其预后     

乳腺浸润性导管癌中Hepsin的表达及临床病理意义

目的：探讨Hepsin蛋白表达与乳腺浸润性导管癌临床病理相关性。方法采用免疫印迹和免疫组化方法检测正常乳腺8例和乳腺浸润性导管癌139例中Hepsin蛋白表达情况。采用统计学方法分析Hepsin蛋白表达与临床病理的关系。结果免疫印迹和免疫组化方法检测所有乳腺浸润性导管癌样本中98例（70．5％）Hepsin高表达,正常乳腺组织未见明显表达或弱表达。病理分期越晚,Hepsin蛋白表达程度越高;组织分化程度与Hepsin蛋白表达程度呈正相关。结论 Hepsin蛋白表达与乳腺浸润性导管癌进展密切相关,Hepsin蛋白表达与临床分期呈正相关,而与组织分化程度负相关。     

乳腺浸润性导管癌中MGMT、XRCC1蛋白表达及意义

目的 探讨乳腺非特殊型浸润性导管癌中MGMT和XRCC1的表达及临床意义.方法 采用S-P免疫组化法检测72例乳腺非特殊型浸润性导管癌石蜡标本、30例乳腺良性病变石蜡标本中MGMT和XRCC1蛋白的表达,分析其表达与临床因素的相关性.结果 72例乳腺癌组织中MGMT阳性表达率为70.8%,低于良性病变组织表达率90.0%（P＜0.05）,且其表达与非特殊型浸润性导管癌的病理分化程度、ER及p53表达相关（P＜0.05）.XRCC1阳性表达率为52.8%,低于良性病变组织表达率83.3%（P＜0.05）,且其表达与非特殊型浸润性导管癌病理分化程度相关（P＜0.05）.结论 DNA修复基因MGMT及XRCC1是临床评估乳腺癌恶性程度、判断预后及制定治疗策略的重要病理指标.     

前列腺癌中p53和p16蛋白的表达及其临床病理学意义

目的 探讨抑癌基因在前列腺癌组织中的表达及临床意义。方法 应用免疫组织化学方法,检测２５例前列腺癌中ｐ５３和ｐ１６基因蛋白产物的表达。结果 ｐ５３蛋白阳性率为３２％（８／２５）,明显高于良性前列腺增生（Ｐ〈０．０５）,并在低分化肿瘤中表达高（Ｐ＝０．０４０３）;在临床分期Ⅲ￣Ⅳ期癌中表达升高（Ｐ〈０．０５）。ｐ１６的失活率４４％（１１／２５）,也是低分化组高于高分化组（Ｐ〈０．０５）。并发现４例低     

p16蛋白在卵巢上皮性浆液性肿瘤中的表达及其临床意义

目的探讨ｐ１６蛋白在正常卵巢和卵巢上皮性浆液性肿瘤中的表达及其临床意义。方法应用免疫组化法检测ｐ１６蛋白在８４例卵巢上皮性浆液性肿瘤组织（良性２０例，交界性１２例，癌５２例）及１２例正常卵巢组织常规石蜡标本的表达。结果ｐ１６蛋白在卵巢浆液性癌中的检出率５０．０％，低于良性浆液性瘤８５．０％及正常卵巢组９１．７％，均Ｐ＜０．０５。ｐ１６蛋白低表达与卵巢浆液性癌的恶性程度高，临床分期晚、肿瘤分化差、癌瘤播散、淋巴结转移及患者预后有关（Ｐ＜０．０１或Ｐ＜０．０５）。结论ｐ１６蛋白作为细胞周期的负性调节子具有抑制细胞增殖的作用。ｐ１６抑癌基因异常导致ｐ１６蛋白合成障碍，可引起细胞无限性生长即恶变。对卵巢浆液性癌ｐ１６蛋白表达的测定，有助于确定肿瘤的恶性度，判断其癌瘤播散、转移的能力，并估计患者的预后     

胰腺癌细胞凋亡相关基因的表达及相互关系

本文采用免疫组化ＳＰ法检测４８例胰腺癌和１５例胰腺癌转移灶石蜡标本中ｂｃｌ－２、ｐ５３、ｃ－ｍｙｃ的表达。结果显示：ｂｃｌ－２蛋白在胰腺癌组明显高于胰癌转移灶组；无转移的、高分化的胰腺癌明显高于有转移的和低分化胰腺癌。ｐ５３蛋白在胰腺癌组明显高于胰腺良性病变组，胰腺高分化癌明显低于胰腺低分化癌。ｂｃｌ－２和ｐ５３蛋白在胰腺癌组中呈负相关。     

Distribution of myoepithelial cells and basement membrane proteins in the normal breast and in benign and malignant breast diseases

An immunocytochemical method for fixed and paraffin-embedded human breast biopsies is reported for the detection of myoepithelial and epithelial cells using antibodies to myosin and keratin, respectively, and of basement membranes using antibodies to laminin and type IV collagen. Using these markers, myoepithelial cells can be clearly distinguished in the normal breast and in the benign breast diseases sclerosing adenosis, epitheliosis, and fibroadenoma. In sclerosing adenosis, myoepithelial cells form a major cellular component. A stromally derived spindle cell is identified which stains with myosin but not with keratin antibodies (myofibroblast). These cells are seen in one-fifth of the fibroadenomas. Although cells staining with myosin antibodies are seen in the infiltrating component of all 18 carcinomas examined, elongated cells staining with both myosin and keratin antibodies (myoepithelial-like) are seen in only one infiltrating carcinoma where they are interposed at the stromal-epithelial junction of the infiltrating tumor cells. In contrast to the situation in benign breast diseases, mature myoepithelial cells form a very minor component of the majority of infiltrating ductal carcinomas. Basement membrane proteins, laminin, and type IV collagen are present in normal breast, benign breast disease, and grade I infiltrating ductal carcinomas but are absent in carcinomas of grades II and III.     

乳腺癌组织Ezrin表达及其与临床病理特征关系的研究

目的:研究Ezrin表达与乳腺癌临床病理特征的关系。方法:应用组织芯片技术和免疫组织化学方法检测117例浸润性乳腺癌和41例乳腺良性增生中,Ezrin、ER、PR、c-erbB-2、MMP-2及MMP-9的表达。结果:Ezrin在91.89%(34/37)的乳腺良性增生中表达于导管腔缘上皮顶膜,在细胞质内无表达;在76.99%(87/113)的乳腺癌组织中,Ezrin表现为细胞质中弥散表达,未见顶膜表达,两者差异有统计学意义,P=0.000。在乳腺癌组织中,Ezrin表达与肿瘤最大直径、TNM分期、淋巴结转移以及基质细胞中MMP-2和MMP-9的表达呈正相关,P值分别为0.016、0.002、0.036、0.007和0.002;而与年龄、组织学分级、ER、PR、c-erbB-2、肿瘤细胞中MMP-2及MMP-9表达无明显相关性。结论:Ezrin在乳腺良恶性病变中有明显不同的亚细胞定位,其表达情况对乳腺病变性质的判断具有重要价值。高表达Ezrin的乳腺癌具有更高的侵袭性和淋巴结转移能力,在乳腺癌组织中检测Ezrin的表达能为患者的预后和淋巴结转移风险评估提供重要参考。     

用单克隆抗体SC3A检测人乳腺肿物组织中的相关抗原

本文以抗人结肠癌相关抗原的ＭＡｂＳＣ３Ａ为探针，用免疫组化法检测了其相应抗原在１５３例１４种不同组织类型乳腺肿物组织中的表达与分布。结果在８０例９型的乳腺癌中有７８例（９７．５％）呈阳性结合，其中５７例（７１．３％）的反应范围在５０％以上，表明这种相关抗原在乳腺癌中有较高的表达，这对乳腺癌的有关研究具有实际应用意义，此抗原在７３例５种乳腺良性病变中亦有４５．２％的阳性表达，提示其与乳腺癌之间可存在     

Fibrinogen deposition without thrombin generation in primary human breast cancer tissue.

The occurrence and distribution of components of coagulation pathways in situ were determined using immunohistochemical techniques applied to 10 cases of primary carcinoma of the breast, normal breast tissue obtained from two patients undergoing reductive mammoplasty, and three patients with benign breast tumors. Tumor cells stained for factor X and thrombomodulin but not for tissue factor, factor V, factor VII, or factor XIII. Rare nonneoplastic duct epithelial cells stained for thrombomodulin, but these tissues did not otherwise stain for any of these antigens. Macrophages within the tumor stroma stained for tissue factor, factor VII, and factor XIII but not for factor V or factor X. These features of macrophages were the same in malignant and nonmalignant breast tissue. Fibrinogen was present in abundance throughout the connective tissue in breast cancer but not in nonmalignant tissues. By contrast, no staining was observed using fibrin-specific antibodies. These results suggest that an intact coagulation pathway does not exist in breast cancer tissue and that thrombin capable of transforming fibrinogen to fibrin is not generated in significant amounts in this tumor type. While fibrin is not a feature of the connective tissue stroma in breast cancer, it is conceivable that the abundant fibrinogen present in the tumor connective tissue (and factor XIII present in connective tissue macrophages) might contribute to the structural integrity of breast tumor tissues.     

Occurrence of components of fibrinolysis pathways in situ in neoplastic and nonneoplastic human breast tissue

The occurrence and distribution of components of fibrinolysis pathways were determined using immunohistochemical techniques applied to 10 cases of primary carcinoma of the breast, normal breast tissue obtained from two patients undergoing reductive mammoplasty, and three cases of benign breast tumors. Tumor cells stained for urokinase- and tissue-type plasminogen activators, plasminogen activation inhibitor-1, plasminogen, and plasmin-antiplasmin complex neoantigen. The tumor connective tissue stained for fibrinogen and its D fragment plasmin digestion product. By contrast, only occasional nonneoplastic duct epithelial cells stained for urokinase- and tissue-type plasminogen activators and there was little or no staining for the other antigens tested. These results are consistent with the existence of local amplification of expression of enzymatically active plasminogen activators, and particularly of urokinase-type plasminogen activator, in situ in primary breast cancer tissue. These features distinguish malignant from benign breast tissue and may modulate neoplastic progression through an effect on tumor cell proliferation, invasion, and metastatic dissemination.     

Estrogen receptor immunocytochemical assay (ERICA) and laminin detection in 130 breast carcinomas and computerized (Samba 200) multiparametric quantitative analysis on tissue sections

An estrogen receptor immunocytochemical assay (ER-ICA) was applied to 130 malignant breast carcinomas and the results were compared to those of steroid binding assays performed on cytosol extracts of the same tumors. Also laminin (lam) distribution was studied in the same tumors. A semi quantitative analysis and a computerized image analysis system (SAMBA 200 TITN) were used to evaluate the positive ER and lam immunostaining. Positive ER immunostaining was always located in the nucleus of tumor cells and of normal cells in adjacent breast tissue. When immunohistochemical staining was correlated to biochemical assay there was a 88% correlation staining intensity and percentage of positive cells significantly increased (p less than 0.01) with cytosolic ER levels. Lam was observed within vascular and epithelial basement membranes (BMs). Lam staining displayed a continuous linear pattern in intraductal carcinomas or was heterogeneously distributed with a discontinuous linear pattern in invasive carcinomas. No intracellular lam staining was detected. In tumors, laminin immunostaining revealed often multilayered BMs and abnormal multilayered BMs in blood vessels in the tumor stroma. These results indicated that (ER-ICA) is to date the most reliable histochemical method for ER detection and correlated in 88% of the cases with ER biochemical assay ER-ICA provides additional information for heterogeneous ER distribution within tumors ER-ICA as a qualitative method is unable to replace the quantitative ER determination obtained with biochemical assay ER-ICA based on ER antigenic site detection is complementary to biochemical assay based on ER functional site determination laminin immunostaing constitutes a new approach to the heterogeneous BM changes occurring in carcinomas, and permits a better understanding of cell diffusion processes and of stroma-tumor cells interactions: the consistent extracellular lam distribution in contact with the stroma, indicates that the latter plays an important role in the assembly of BM components the SAMBA 200 permits a reliable accurate evaluation of the percentage of the immunostained cells and surfaces.     

CORRELATION BETWEEN LAMININ AND CATHEPSIN D EXPRESSIONS IN BREAST CARCINOMA

Objective： Laminin is a major glycoprotein component of basement membrance which is an important barrier to tumor cells which must be breeched before metastatic spread can occur. Proteolytic enzymes play an important role in mediating the passage of cancer cells through the basement membrane （BM） and extracellular matrix. We compared the patterns of laminin and cathepsin D （CD） expressions in a range of benign and malignant breast lesions to better understand the process of tumor progression. Methods： One hundred and sixty-two cases of breast samples comprising 18 fibroadeomas, 22 cases of fibrocystic disease, 96 cases of invasive ductal carcinoma and 26 carcinomas with intraductal components were evaluated for laminin and cathepsin D expressions by immunohistocbemical staining. Results： The prevalence of CD positivity in both neoplastic and stromal cell components were significantly higher in higher histological grade tumors compared to lower grades （P〈0.001）. Various severity of BM disruption correlated with histological grade of the carcinomas （P〈0.001）. There was a negative correlation between the laminin expression and CD presence. Conclusion： In the process of cancer cell invasion and metastasis, the basement membrane is disrupted by proteinase secreted by cancer cells, especially by stroma cells of cancer.     

VEGF-C在乳腺癌组织中的表达及临床意义

目的检测血管内皮生长因子C(VEGF-C)在乳腺癌、乳腺良性肿瘤及正常乳腺组织中的表达,探讨VEGF-C与乳腺癌淋巴结转移及预后的关系。方法选用10例正常乳腺组织、30例乳腺良性肿瘤及60例乳腺恶性肿瘤标本,以免疫组织化学及半定量反转录PCR(RT-PCR)检测VEGF-C的表达。结果在乳腺恶性肿瘤组织中,VEGF-C阳性表达率为46.7%,明显高于正常乳腺(10.0%)和乳腺良性肿瘤组织(13.3%)(P<0.01);乳腺癌中VEGF-C的阳性表达率为48.2%,明显高于乳腺叶状囊肉瘤(25.0%)(P<0.01);在伴有淋巴结转移的乳腺癌组织中,VEGF-C的阳性表达率(55.0%)明显高于没有淋巴结转移的乳腺癌组织(30.0%)(P<0.01)。结论VEGF-C在乳腺癌组织中表达明显增高,并与乳腺癌的淋巴结转移显著相关,可作为乳腺癌转移、复发的预测指标之一。     

Detection of the plasmin system in human mammary pathology using immunofluorescence

The presence and localization of the plasmin system components urokinase (UPA), tissue type plasminogen activator (TPA), plasminogen (PG), a neoantigen expressed by the plasmin-alpha 2-antiplasmin complex, and plasmin inhibitors alpha 2-antiplasmin (AP) and alpha 2-macroglobulin (MG) have been tested by immunofluorescence on sections of 11 benign and 40 malignant lesions of the breast in an attempt to apply a morphological approach to the problem of tumor invasion in vivo. In benign lesions, TPA was seen in secretions of mammary glands and MG was seen in edematous zones. In one involuting lactating adenoma, UPA, TPA, PG, PAP, and AP were associated with glandular cells. UPA was detected in 11 carcinomas, TPA in 22, PG in 31, PAP in 12, AP in 23, and MG in all 40. All these components were essentially present in invasive territories, with a cellular labeling for UPA and TPA and a fluorescent staining frequently at the periphery of tumoral foci for PG and PAP. AP was more closely associated with cancer cells than MG, which was present in the stroma. Intraductal proliferations were rarely positive and there was no correlation between the localization of PG and the distribution of a basement membrane glycoprotein laminin. These data argue strongly for the involvement of the plasmin system in the infiltrating process of the stroma. This system seems to play a limited role in the breakdown of basement membrane in breast carcinomas in vivo.     

Immunohistologic c-myc protein in benign breast disease and cancer

We have studied the histopathology and differential distribution of the c-myc protein (Myc) in human breast tissues including 17 cases of infiltrating mammary carcinoma, 4 cases of fibroadenoma, 5 cases with fibrocystic changes, and 1 case of reduction mammoplasty (as a control). Using a sensitive immunohistochemical method on frozen tissue sections, both a rabbit polyclonal anti-c-myc antibody and a mouse monoclonal anti-c-myc antibody, H51C116, produced high levels of Myc staining in the nuclei of epithelial cells of infiltrating mammary carcinomas (30-90% of cells stained). In contrast, the nuclei of epithelial cells of fibroadenomas, and breast tissues with fibrocystic changes stained infrequently. We studied benign tissue surrounding the tumors in four cases; three were essentially negative, and one showed nuclear epithelial cell staining throughout the lobules. Sixteen of the tumors were examined in parallel, using formalin-fixed, paraffin-embedded samples. Immunohistological procedures for Myc produced uniform, intense epithelial cell cytoplasmic staining (8 cases); light epithelial cell cytoplasmic staining (5 cases) or were unstained (3 cases). We argue that the differences between frozen and paraffin sections are incompatible with the notion of simple displacement of nuclear Myc to the cytoplasm during fixation. Elevated levels of nuclear Myc in tumor cells and subsets of benign tissue are consistent with a role for Myc in mammary cell proliferation and tumorigenesis.