Superoxide dismutases in relation to the overall survival of colorectal cancer patients.

Reactive oxygen metabolites are implicated in the initiation and promotion of cancer. In addition, oxidant scavengers, such as manganese--(Mn-SOD) and copper/zinc--superoxide dismutase (Cu/Zn-SOD), are thought to contribute to colorectal cancer treatment response. In the present study, the prognostic significance of the Mn- and Cu/Zn-SOD antigen content of normal mucosa and carcinomas of 163 patients with colorectal cancer was evaluated in comparison with major clinicopathological parameters, with respect to the 5-year overall survival. The Mn-SOD content of carcinomas was found to be significantly higher than that of normal mucosa, whereas there was no difference in the Cu/Zn-SOD content between the normal mucosa and carcinomas. No association was demonstrable between the Mn-SOD and Cu/Zn-SOD content of the tissues and the assessed clinicopathological parameters (gender, age, localization, differentiation grade, diameter and Dukes'' stage), with the exception of the Cu/Zn-SOD and the differentiation grade of the carcinomas. Univariate analysis showed that a high Mn-SOD content of carcinomas was associated with a poor 5-year overall survival of the patients with colorectal cancer. Multivariate analysis including all clinicopathological parameters revealed that this Mn-SOD parameter was prognostically independent. The Mn- and Cu/Zn-SOD content of normal mucosa and the Cu/Zn-SOD content of carcinomas were not associated with the overall survival of the patients. In conclusion, this study demonstrates that for patients with colorectal cancer the Mn-SOD content of colorectal carcinomas has a significant prognostic value that is independent from major clinicopathological parameters, including Dukes'' stage.     

Expression of Cu,Zn-SOD, Mn-SOD and GST-pi in oral cancer treated with preoperative radiation therapy

Radical scavengers play an important role in cancer cells defending themselves against free radicals which occur with irradiation. SOD (Cu,Zn, Mn-) and GST-pi are radical scavengers with an effect on radiation therapy. We investigated the correlation between radiation effects and expression of Cu,Zn-, Mn-SOD and GST-pi in 34 cases of oral cancer, treated with preoperative radiation therapy. In this study, 22 cases out of 34 were classified as effective and 12 cases as non-effective. Expression of Cu,Zn, Mn-SOD and GST-pi were observed in 13 (38.2%), 10 (29.4%) and 20 (58.8%) cases, respectively. Regarding the value of radiation sensitivity from expression of these proteins in the biopsy samples, no significant correlation was found between those expressions and histological effectiveness of preoperative radiation therapy. But interestingly, in 11 out of 12 of the non-effective cases, strong staining of Cu, Zn-SOD and GST-pi were shown at the residual cancer cells after preoperative radiation therapy. These results suggested that the expression of SOD (Cu,Zn-, and Mn-) and GST-pi may be not useful markers for predicting the effects of radiation therapy. However, Cu, Zn-SOD and GST-pi were increased by irradiation and may play an important role in radiation resistance and cancer cell regeneration after radiation therapy.     

Beta-chemokine, macrophage inflammatory protein-1beta (MIP-1beta), is highly expressed in diffuse type human gastric cancers

Chemokines have been shown to be expressed in some malignant or precancerous tissues. However, the role of these chemokines on tumor development or progression is not clear. The expression patterns of chemokines in gastric cancer tissues were examined in 86 surgically resected samples using immunohistochemistry. Macrophage inflammatory protein-1beta (MIP-1beta) was clearly detected in many gastric carcinoma cells. In most of the differentiated carcinomas, intracellular localization of MIP-1beta was detected in more than 5% of cancer cells, although the percentages of MIP-1beta-positive cells differed among each sample. Undifferentiated carcinomas showed contrasted staining pattern between solid type and non-solid (diffuse) type. MIP-1beta was totally absent in all the poorly differentiated carcinomas with solid type growth pattern (por1). In contrast, MIP-1beta was highly expressed in all of the non-solid type of poorly differentiated carcinoma (por2) and signet-ring cell carcinoma samples. In particular, MIP-1beta was strongly stained in carcinoma cells at the front of invasive lesions. In 43 diffuse type undifferentiated cancers, tumors with high expression of MIP-1beta exhibited significantly more lymph node metastasis. Our results suggest a possibility that MIP-1beta may be related to the scattering and invasion step of gastric carcinoma cells with undifferentiated phenotype.     

Stainability of lung cancer cells with Leu-7 and OKT-9 monoclonal antibodies

One hundred and one cases of lung cancer were immunohistochemically studied for stainability with Leu-7 (anti-myelin fiber associated glycoprotein) and OKT-9 (anti-transferrin receptor) monoclonal antibodies. All small cell carcinomas and carcinoid tumors were positively stained by Leu-7, and 22 of 41 differentiated adenocarcinomas were also positively stained, especially well-differentiated Clara cell-type adenocarcinoma, (11/14 cases). However, only one of 26 squamous cell carcinomas, one of six large cell carcinomas, one of six adenosquamous carcinomas and none of 16 poorly differentiated adenocarcinomas were stained by Leu-7. On the other hand, all squamous cell carcinomas (26/26 cases), 10 of the 16 poorly differentiated adenocarcinomas, four of the six large cell carcinomas showed positive membranous staining with OKT-9. However, only one of 41 differentiated adenocarcinomas and no small cell carcinomas nor carcinoid tumors were stained by OKT-9. The stainability of lung cancer by these antibodies is discussed.     

Study of the antioxidant enzymes in human brain tumors

Summary The activities of antioxidant enzymes i.e. Cu, Zn-SOD, Mn-SOD, CAT, and GSH-Px in the normal brain and brain tumors, as well as the two varieties of SOD in the mitochondria were examined and correlated to the histopathological diagnosis and the degree of malignancy of tumors. It was found that these scavenging enzymes of oxygen free radicals were expressed with great regularity in brain tumors. Both Cu, Zn-SOD and Mn-SOD were decreased in descending order in meningiomas, low grade astrocytomas, high grade astrocytomas and medulloblastomas. Furthermore, the reduction of Mn-SOD in mitochondria was proportionate to that of the whole tissues. While in contrast to the SODS, the CAT levels were significantly increased in ascending order in high grade astrocytomas, low grade astrocytomas and meningiomas. GSH-Px increased in meningiomas but not in gliomasAbbreviations SOD superoxide dismutase - Cu, Zn-SOD copper and zinc containing superoxide dismutase - Mn-SOD manganese containing superoxide dismutase - CAT catalase - GSH-Px glutathione peroxidase - GSH reduced glutathione - GSSG oxidized glutathione - RBC red blood cell     

Immunohistochemical localization of urokinase-type plasminogen activator,type-1 plasminogen-activator inhibitor,urokinase receptor and α2-macroglobulin receptor in human breast carcinomas

We have investigated the localization of urokinase-type plasminogen activator (u-PA), type-1 plasminogen-activator inhibitor (PAI-1), u-PA receptor (u-PAR) and α₂-macroglobulin-receptor/low-density-lipoprotein-receptor-related protein (α₂MR/LRP) in human breast tumors by immunohistochemical methods. Frozen sections of 133 primary breast carcinomas, 6 ductal carcinomas in situ and 33 lymph-node metastases were stained with monoclonal antibodies. Formalin-fixed sections of 15 primary tumors and 2 lymph-node metastases were stained with polyclonal antibodies. In primary tumors, u-PA and PAI-1 immunoreactivities were intense in macrophages and mast cells, and moderate in benign and malignant epithelial cells as well as in myofibroblasts and endothelial cells. A sub-group of poorly differentiated tumors showed particularly strong staining of stromal fibroblasts. u-PA immunoreactivity was also present in lymphocytes. α₂MR/LRP and u-PAR immunoreactivities were intense in macrophages, but apart from these cells, α₂MR/LRP was found only in fibroblasts, and u-PAR only in tumor cells located peripherally in tumor-cell clusters and glands and some myofibroblasts in the adjacent stroma. Lymph-node metastases showed staining for u-PA and PAI-1 both of cancer cells and of stromal fibroblasts, also staining for u-PA of lymphocytes. Similarly to some of the poorly differentiated primary tumors, approximately half of the metastases showed very strong staining of stromal fibroblasts, and extracts of these metastases had higher u-PA and PAI-1 levels, as determined by ELISA, than extracts of metastases without this staining pattern. α₂MR/LRP was present only in fibroblasts and u-PAR only in some tumor cells. The presence of u-PA, PAI-1, α₂MR/LRP and u-PAR was controlled biochemically by immunoblotting analyses, ligand-blotting analyses, and direct and reverse zymography. The spatial distribution and the variation in concentration of the various components of the plasminogen-activation system point to a complex, multifunctional role for the 4 proteins in and/or during the development and spread of breast cancer. © 1996 Wiley-Liss, Inc.     

Expression of nm23-h1 gene in squamous-cell carcinoma of the cervix correlates with 5-year survival

We have examined nm23-H1 protein levels in human squamous cell carcinoma of the cervix by immunohistochemistry and correlated detectable nm23-H1 with patient survival. Normal cervical tissue samples were consistently strongly immunoreactive. In contrast, cervical carcinomas stained poorly, if at all. Well differentiated carcinomas stained more frequently for nm23-H1 protein than poorly or undifferentiated carcinomas. Our results clearly demonstrated a significant correlation between absence of nm23-H1 protein staining and shorter survival of patients with cervical cancer. Our results suggest that nm23 may serve as a potentially valuable prognostic marker for this type of tumor and to our knowledge is the first report of this kind for cervical carcinoma.     

Expression of placental alkaline phosphatase in gastric and colorectal cancers. An immunohistochemical study using the prepared monoclonal antibody

The authors developed monoclonal antibodies (MoAb) against human placental alkaline phosphatase (PLAP). Four specific MoAb reacting only with PLAP and two nonspecific MoAb reacting equally with isozymes of alkaline phosphatase (hepatic, intestinal, and placental) were obtained. Immunohistochemical staining with the specific MoAb showed that the cell membrane and cytoplasm of cancer cells were stained in gastric and colorectal carcinoma. The incidence of PLAP positivity was 23% (25 of 107) of all gastric carcinomas. Among gastric carcinomas, the 42% (13 of 31) positivity of highly differentiated carcinoma (papillary adenocarcinoma and well-differentiated tubular adenocarcinoma) was a significantly higher rate than that found in poorly differentiated carcinoma (poorly differentiated adenocarcinoma and signet-ring cell carcinoma, five of 41, 12%). The incidence of PLAP positivity was 11% (four of 35) in colorectal carcinoma. In contrast, gastric adenoma, intestinal metaplasia, and noncancerous tissue adjacent to cancer did not show staining. These results indicated that expression of PLAP was apt to occur in more highly differentiated gastric carcinoma and was highly specific for carcinoma in the gastrointestinal tract, although its incidence was not high.     

High copper and iron levels and expression of Mn-superoxide dismutase in mutant rats displaying hereditary hepatitis and hepatoma (LEC rats)

The LEC rat is a mutant strain displaying hereditary hepatitisand hepatoma. We established enzyme-linked immunosorbent assaysof Cu, Zn- and Mn-superoxide dismutase (Cu,Zn- and Mn-SOD) andmeasured the levels of both SODs in various organs of LEC andWistar rats. Mn-SOD concentrations were higher in LEC rats thanin Wistar rats in most tissues. Cu,Zn-SOD levels of liver, kidneyand intestine were lower in LEC rats than in Wistar rats. Atomicabsorbtion techniques indicated that in addition to high Cuconcentrations as previously reported, LEC rat livers containedhigh Fe concentrations relative to those in Wistar rat livers.These data suggest that increased concentrations of Fe and Cuand decreased levels of Cu,Zn-SOD may facilitate the Fentonreaction to produce hydroxyl radicals in the tissues of theLEC rat. To compensate for the decreased scavenging effectsdue to low levels of Cu,Zn-SOD, an adaptive increase of Mn-SODmay occur in the process of hepatitis and hepatocarcinogenesisin LEC rats.     

金属硫蛋白和超氧化物歧化酶在颊鳞状细胞癌组织中的表达

背景与目的探讨金属硫蛋白(Metallothionein,MT)及铜/锌超氧歧化酶(Cu/Zn Superoxide dismutase,Cu/Zn-SOD)在颊粘膜鳞状细胞癌的表达及其意义。材料与方法对8例颊癌MT和Cu/Zn-SOD采用免疫组化的方法,观察两者在颊癌中表达的定位,并对MT和CuZn-SOD表达进行定量分析。结果在高分化鳞癌,MT和CuZn-SOD主要表达在癌巢周边,癌巢中心及间质组织表达较少或不表达,在低分化鳞癌则散在分布。在8例颊癌标本中,MT和Cu/Zn-SOD表达差异无统计学意义(P=0.554)。结论MT和Cu/Zn-SOD主要表达在分化较差、生长活跃的癌细胞,两者相类似的表达对肿瘤的发生发展和预后关系值得进一步探讨。     

Immunohistochemical determination of estrogen and progesterone receptors in human breast carcinoma. Correlation with histopathology and DNA flow cytometry

Human breast carcinomas (n = 232) were evaluated for estrogen and progesterone receptors (ER, PR) by immunohistochemical study and by cytosol steroid-binding assay (n = 185). The staining was scored (histoscore) by estimates of relative nuclear staining intensity and the percentage of positively stained carcinoma cells. Of the invasive ductal carcinomas 72% were ER-positive and 55% were PR-positive. The invasive lobular, intraductal, tubular, and mucinous carcinomas were the most frequent ER-positive tumor types, whereas comedo and medullary carcinomas only rarely contained ER. Progesterone receptor was most frequently present in intraductal, tubular, and mucinous carcinomas. Better differentiated tumors with lower histologic grade were significantly associated with high prevalence of immunohistochemically determined ER and PR (P less than 0.0001). Proliferative cell fraction, determined by DNA flow cytometric study (n = 63), was inversely related to ER (P = 0.03) and PR (P = 0.05) status. Aneuploidy was independent of ER or PR content.     

Association of progression and reduced expression of VLA-2 in gastric cancer

We investigated the expression of VLA-2 in gastric cancers by immunohistochemistry using anti-integrin alpha 2 and beta 1 antibodies and the data were compared with the pathological findings of each gastric cancer. The specimens were stained with an immunohistological technique for integrin alpha 2 and beta 1 subunits. Tumors, simultaneously expressing both integrin alpha 2 and beta 1 subunits were defined as positive for VLA-2. Tumors expressing either subunits of integrin alpha 2 or beta 1 or those showing reduced expression of both subunits were defined as VLA-2 negative tumors. In the 77 primary tumors, 55 (71%) were VLA-2 positive. 38 (90%) of 42 tumors showing differentiated type including tubular adenocarcinoma and papillary adenocarcinoma expressed VLA-2, whereas 19 (55%) out of 35 undifferentiated type of cancers including poorly differentiated adenocarcinoma, mucinous carcinoma and signet ring cell carcinoma stained for VLA-2. In the undifferentiated type of cancers, VLA-2 negative tumors had a significantly higher incidence of vessel invasion than VLA-2 positive ones (p<0.05). VLA-2 negative tumors showed a tendency to peritoneal dissemination, lymph node metastases, lymphatic invasion or invasion beyond the subserosal layer. In the specimens of peritoneal dissemination, VLA-2 expression rate was found in 56% (9/16), with a higher expression rate than that of primary lesions. These data indicate that reduced expression of VLA-2 may strongly associate with vessel invasion especially in the undifferentiated type of adenocarcinoma of the stomach.     

Time trends for small gastric cancer in Japan

Background. Gastric cancer rates in Japan have been declining since the 1970s. The rate of differentiated carcinomas has decreased and that of undifferentiated carcinomas has increased. However, little is known about the time trends of small gastric cancer. The aim of this study was to investigate the trends of small gastric cancer over time in Japan. Methods. We reviewed cases of small gastric cancer (less than 20 mm in diameter) in two groups of patients who entered the age range of 55-to-67 years 14 years apart: patients in cohort 1 (n = 66) were born between 1899 and 1912, and those in cohort 2 (n = 66) were born between 1926 and 1936. Between-group comparisons were made for macroscopic, microscopic, and histochemical findings. Mucin histochemical analysis was used to investigate gastric and nongastric phenotypes. Helicobacter pylori was also investigated by immunohistochemistry. Results. There were significant decreases in the incidence of elevated carcinoma (20% in cohort 1 vs 6% in cohort 2; P < 0.05) and papillary adenocarcinoma (11% vs 2%; P < 0.05). The incidence of flat carcinomas was significantly increased (3% vs 15%; P < 0.05). The incidence of tumors surrounded by fundic gland mucosa increased (20% vs 29%), whereas that of tumors surrounded by intestinal metaplastic mucosa decreased (52% vs 41%). The rate of H. pylori infection in mucosa surrounding tumors was the same in both groups (35%). The incidence of tubular adenocarcinoma with gastric-type mucin was higher in cohort 2 (64%) than in cohort 1 (51%). Conclusion. The rate of tubular adenocarcinomas containing gastric type mucin has increased over time. These tumors had a tendency to develop in the fundic gland mucosa and to show less intestinal metaplasia. The H. pylori infection rate was unrelated to this time trend. In advanced gastric cancer, the differentiated carcinoma rate has decreased; however, in small gastric cancer, the rate of tubular adenocarcinoma containing gastric type mucin has increased. This suggests that tubular adenocarcinoma with gastric type mucin changes into poorly differentiated adenocarcinoma as tumors grow to advanced stages. Received: April 24, 2000 / Accepted: August 1, 2000     

Altered expression of E-cadherin in gastric cancer tissues and carcinomatous fluid.

Expression of E-cadherin in 21 patients with various histological types of gastric carcinomas was studied by immunoperoxidase staining. Intercellular boundaries of almost all cancer cells in well and moderately differentiated adenocarcinomas stained as deeply for E-cadherin as normal gastric mucosa. However, singly infiltrating cells of those histological types were poorly stained. In poorly differentiated adenocarcinomas, cancer cells forming clusters stained lightly and those infiltrating singly stained even less. In signet ring cell carcinomas, hardly any staining was observed. In each histological type, the staining patterns and intensity at different layers of the gastric wall, were essentially the same. Cancer cells from carcinomatous ascites of gastric adenocarcinomas and pancreatic adenocarcinomas, and those from pleural effusion of lung adenocarcinomas were also studied by immunofluorescence staining. Of 11 specimens, ten were negative and only one from a lung adenocarcinomas was positively stained. By phase-contrast microscopic observations, none of these cancer cells including those from the lung adenocarcinomas, formed obvious cell-cell contacts. Cell aggregation assays confirmed the above results. The molecular weight of E-cadherin of cancer cells of lung adenocarcinomas was less than intact E-cadherin as revealed by Western blot analysis. These results suggest that depressed expression and/or impaired function of E-cadherin in cancer cells, facilitates their liberation from primary sites to infiltrate freely into tissue or fluid.     

Tissue distribution of an epithelial and tumor-associated antigen recognized by monoclonal antibody F36/22

Murine monoclonal antibody F36/22 was derived by immunizing BALB/c mice with human breast cancer cells. This antibody reacts with an antigen located both on differentiated mammary ductal epithelia and on breast carcinomas, as examined by indirect immunoperoxidase techniques. Although the expression of this antigen correlated with estrogen receptor levels of breast tumors, antibody F36/22 did not directly react with estrophilin. In contrast to the expression of classical differentiation antigens, this antigen was found in a high percentage of poorly differentiated carcinomas of the breast. Staining intensities were similar for well- and poorly differentiated tumors; thus, antigen expression was not related to tumor grade. Intratumoral heterogeneity of antigen expression was observed in the majority of tumors. Since a subset (64 of 80) of the breast carcinomas examined have expressed the antigen, McAb 36/22 was of use for the immunological subclassification of tumors which were indistinguishable by conventional histopathological staining techniques. The antigen was also present on other adenocarcinomas (ovary, colon, stomach, pancreas, and prostate); however, these tumors usually exhibited reduced staining intensity compared with that observed in breast cancer. The normal counterpart tissues at these histotypes contained no detectable levels of the antigen, and increased expression of the antigen was associated with tumorigenesis at these sites. Tumors of mesenchymal origin and carcinomas other than adenocarcinomas exhibited undetectable levels of the antigen. Therefore, depending on the organ site, McAb F36/22 recognizes an epithelial and/or tumor-associated antigen.     

Histiocytoid carcinoma: a variant of breast cancer

The history of a patient presenting with metachronous bilateral breast cancer displaying histiocytoid features is reviewed. Although regional metastases were noted, this patient has not demonstrated an aggressive systemic disease pattern. In the past, histiocytoid breast cancer has been classified as either a lipid-rich carcinoma or as a variant of lobular carcinoma. However, histiocytoid carcinoma should be considered a distinct entity. Unlike the lipid-rich carcinomas, this tumor stained strongly for mucin. Immunoperoxidase staining indicated strong positively for CEA and negative staining for alpha-lactalbumin. There is suggestive evidence of a relationship between histiocytoid breast carcinoma and breast cancers of apocrine origin. Controversy remains and further evaluation is needed to elucidate the histiogenesis and biological potential of this neoplasm.     

Copper-transporting P-Type Adenosine Triphosphatase (ATP7B) Is Expressed in Human Breast Carcinoma

This is the first report to show that a copper-transporting P-type adenosine triphosphatase, ATP7B, is expressed in certain breast carcinomas, and a priori knowledge of its expression is important for the choice of therapy. We investigated the hypothesis that ATP7B, which was shown to be associated with cisplatin resistance in vitro , is expressed in certain breast carcinomas. To test this hypothesis, ATP7B expression and protein level were examined in 41 breast carcinomas using RT-PCR and immunohistochemistry. ATP7B gene/protein could be detected in 22.0% (9/41) of breast carcinomas and ATP7B gene expression was correlated well with the protein expression. In nine ATP7B-positive tumors, adjacent normal breast tissue was similarly analyzed, revealing that ATP7B is upregulated in breast carcinoma. ATP7B gene expression in poorly differentiated carcinoma was significantly higher than that in well-/moderately differentiated carcinoma ( P =0.012). Furthermore, we found no association between the ATP7B gene/protein expression and that of MDR1, MRP1, LRP and BCRP. These findings suggested that ATP7B gene expression might be a chemoresistance marker for cisplatin in patients with poorly differentiated breast carcinoma.     

免疫标志物及AB／PAS黏液染色在子宫颈癌分型中的应用研究

目的：探讨免疫标志物细胞角蛋白7（cytokeratin 7,CK7）、低分子质量细胞角蛋白CAM5．2、癌胚抗原（carcino-embryonic antigen,CEA）、癌基因p63、高分子质量细胞角蛋白34βE12及阿利新蓝-过碘酸雪夫氏（alcian blue-periodic acid schiff,AB／PAS）黏液染色在子宫颈癌分型中的意义。方法采用免疫组织化学EnVision法检测CK7、CAM5．2、CEA、p63、34βE12在59例子宫颈癌中的表达,结合AB／PAS黏液染色,分析上述免疫标志物及特殊染色与子宫颈癌分型的关系。结果 CK7、CAM5．2、CEA在子宫颈腺癌（6例）、腺鳞癌（4例）的腺癌细胞质中均有100．0％的表达,低分化鳞癌（34例）中CK7、CAM5．2阳性率分别为52．9％、32．4％,而中及高分化鳞癌（15例）中不表达。CEA在鳞癌中有不同程度表达,中及高分化鳞癌仅在角化区域表达,低分化鳞癌中阳性率44．1％;p63在腺癌中不表达,在鳞癌中100．0％表达;34βE12在腺癌及鳞癌中均有100．0％表达,鳞癌中强度高,腺癌中较弱;AB／PAS在子宫颈腺癌、腺鳞癌、低分化鳞癌中发生率分别为83．3％（5／6）、75．0％（3／4）、5．9％（2／34）,但表达程度很弱。结论 CK7、CAM5．2、p63、34βE12联合检测对子宫颈癌的分型有重要意义。AB／PAS黏液染色对子宫颈癌有一定的鉴别作用。