Recognition of N-glycosidic carbohydrates on esophageal carcinoma cells by macrophage cell line THP-1.

Cell-to-cell contact between macrophages and tumor cells is an important initial reaction in a host defense mechanism against tumor cells. The authors have studied cell surface components of human esophageal carcinoma cells recognized by macrophages. Superoxide release from THP-1 cells, a human macrophage cell line, was analyzed in their interaction with a battery of human squamous cell carcinoma cell lines (TE) originated from esophageal cancer patients. The macrophage-triggering ability of TE 1 cell line, a high stimulant, was reduced after treatment with trypsin or tunicamycin, an inhibitor of N-glycosidic glycosylation. Addition of monosaccharides was efficient in competitive inhibition of these cellular interaction. Moreover, con-A-resistant mutation of TE 1 cells was found to reduce their macrophage-triggering ability, associated with increase of L-PHA-binding capacity, suggesting substitution to the GlcNAc beta(1----6)-linked lactosamine antenna in N-glycosidic carbohydrates. These findings suggest that terminal residues of N-glycosidic carbohydrates on some esophageal carcinoma cells may contribute to the recognition sites of macrophages.

Expression of ras oncogene p21 protein in esophageal squamous cell carcinoma.

In order to investigate the value of ras oncogene expression as a prognostic indicator in esophageal squamous cell carcinoma, we evaluated the level of ras oncogene protein product (p21) in 52 specimens resected between 1977 and 1986. All patients were followed until death or for at least 2 years. Pathology slides and archival paraffin blocks were retrieved for confirmation of the original diagnosis, study of histopathologic features, and measurement of p21 content. P21 titers were obtained using the RAP-5 monoclonal antibody in a semiquantitative immunohistochemical assay. Titer was expressed as the highest dilution of antibody giving definitive staining using the avidin-biotin peroxidase method. Ras oncogene was expressed in 88.5% of the specimens. We did not find a significant correlation between ras expression and any of a variety of clinical and histopathologic prognostic parameters. Although patients' median survival after resection of specimens with ras oncogene expression was less than half the median survival after removal of tumors without such expression, this difference was not statistically significant. Further prospective investigations are needed to assess the role of ras oncogene evaluation in clinical practice.

Esophageal carcinoma-associated proteins detected by two-dimensional polyacrylamide gel electrophoresis.

Patterns of proteins of five surgically resected esophageal carcinomas were studied by two-dimensional polyacrylamide gel electrophoresis with silver staining. The samples of normal esophageal mucosa and esophageal carcinoma from the same patient were compared. Each gel had ca. 300 protein spots and had a similar pattern of proteins. Four spots were observed in all of the esophageal carcinomas that were not present in any of the normal mucosae. The molecular weights and isoelectric points were 46,000 and 5.3, 46,000 and 5.2, 36,000 and 4.7 and 33,000 and 5.1, respectively. One spot was observed in all of the normal mucosae but not in any of the esophageal carcinomas. Its molecular weight and isoelectric point were 27,000 and 5.3, respectively.

[Studies on DNA content in ploidy patterns of esophageal cancer].

The present study was undertaken to evaluate the nuclear DNA content of esophageal cancer cells consequent on the process of growth and progression of the cancer. In experimental animal studies, 47 esophageal cancers were induced in male Wistar rats by oral administration of N-amyl-N-methylnitrosamine (AMN) and were analysed in the study of ploidy patterns. The study was also carried out to determine the DNA content in the ploidy patterns in man. Primary tumors associated 421 metastatic lymph nodes in the 62 patients with thoracic esophageal cancer were subjected for the study of ploidy patterns. The nuclear DNA content was determined by means of flow cytometry. In the study of the experimentally-induced esophageal cancer in rats, aneuploidy was found in 18% at a depth of submucosa, 30% at proprial muscle, 59% at adventitia, and in 50% at a depth of neighboring structures, respectively. Clinically in man, the incidence of DNA diploidy and aneuploidy in the 62 primary cancers was 56% and 44%, respectively. In the 421 metastatic lymph nodes, diploid was found in 73% and aneuploid in 11%, while the combination of diploid and aneuploid was observed in 16%. Difference in the DNA index (DI) between the primary cancers and metastatic lymph nodes was found in 29 cases (46.8%), and the difference increased with progression of the cancer. Two hundred and ninety seven metastatic lymph nodes of 29 cases were subdivided into 4 groups based on the extent of the cancerous nests, and the DI value was found to be increased in proportion to the extension. With the results, the DI value of esophageal cancer appeared to be changed dependently by the variation of cell populations in the cancer or in the DNA content of the cancer cells.

Small cell carcinoma of the oesophagus: a clinical and immunohistopathological review.

Primary small cell carcinoma of the oesophagus (SCCO), histologically indistinguishable from its counterpart of the lung, is a rare tumour. Less than 100 cases are reported. A review of 558 consecutive patients with oesophageal carcinomas referred to our department revealed seven cases. These were studied and compared to a survey of 80 cases collected from 24 reports. The present results, as well as the survey, indicate a poor prognosis with rapid and widespread dissemination, and that death is attributed to distant metastases rather than local failure. Freedom from local symptoms was achieved in all seven patients, regardless of therapy modalities employed. A complete response of the primary lesion was observed in three patients after chemo- and subsequent radiotherapy. According to these findings the most suitable treatment approach seems to be the same as for small cell carcinoma of the lung. A detailed immunohistochemical analysis revealed more characteristics similar to small cell carcinoma of the lung than that of the skin, viz 'Merkel cell carcinoma'.

[Study on DNA content and nucleus/cytoplasm ratio of esophageal epithelium during malignant change].

DNA content and nucleus/cytoplasm ratio were assayed in the paracancerous mucosa from 37 cases of esophageal carcinoma by microspectrophotometry (Leitz MPV3). The histopathological diagnosis was made at the same area. In normal epithelium and simple hyperplasia, the relative DNA content was within the range of 2C-4C; cells with DNA content of 4C or greater than 4C were found mostly in invasive carcinoma. There were two different types of DNA content histogram observed in atypical hyperplasia and carcinoma in situ. One type was similar to simple hyperplasia and the other type was similar to invasive carcinoma. These results indicated that two different malignant features might be obtained under the same histopathological diagnosis. The nucleus/cytoplasm ratio in simple hyperplasia was close to that of normal mucosa. In atypical hyperplasia and carcinoma in situ, especially in the middle and superficial layers of the epithelium, the nucleus/cytoplasm ratio increased markedly, which indicated that the cells in these layers were poorly differentiated.

[The significance of determination of urinary trypsin inhibitor related antigen in patients with cancer of the digestive organs].

A highly sensitive enzyme immunoassay was developed for the determination of urinary trypsin inhibitor related antigen (UTIRA) in plasma, urine and cancer tissues, and we investigated its significance as a tumor marker in patients with cancer of the digestive organs. Variations in UTIRA in plasma and urine were not associated with the age of the volunteers, but in urine, high average was observed in male volunteers. UTIRA levels in the urine of the patients with esophageal, stomach and colorectal cancer increased as the stage progressed, except in the patients with liver cancer. And those levels in the urine significantly decreased after a curative operation, but did not decrease after a non-curative operation. UTIRA levels in stomach cancer tissues were significantly higher than those in adjacent healthy mucosa, but UTIRA levels were not high in tissues of esophageal, colorectal and primary liver cancers. Therefore, we speculate that the high levels of UTIRA in urine and plasma in patients with cancer of the digestive organs may not be caused by UTIRA released from cancer tissues. The determination of UTIRA may be significant in a broad sense as an index of the cancer stage or recurrence of cancer, particularly of the stomach.

[Cytodynamic evaluation on the relationship between the mode of intracellular DNA content and the prognosis of esophageal cancer patients].

In order to elucidate biological features of esophageal carcinoma influencing the prognosis of patients, intracellular DNA content was determined using a flow-cytometry (FCM) apparatus. DNA ploidy patterns in 112 paraffin-embedded specimens removed at surgery were classified into 4 categories according to the shape of the first peak curve and the presence of the second peak curve. Type A: with symmetric single peak curve. (euploidy type) Type B: with asymmetric single peak curve. Type C: with the second small peak curve appearing on the right side of the first peak curve. Type D: with two obvious peak curves. Type B, C and D were grossly included in aneuploidy type, because of the presence of abnormal stem line. These DNA ploidy patterns were well correlated with histologic types. There were, however no significant differences in the relationship of DNA ploidy patterns and clinical stages. Four-year survival rate of the patients with euploidy pattern was 65.5%, and that with aneuploidy pattern was 15.7%, indicating a significant difference in survival between patients with the former pattern and those with the latter. The type of DNA ploidy was considered as one of the important prognostic indicators in patients with esophageal carcinoma independent of any other pathologic factors.

Screening test for hormone sensitivity by the autoradiographic method using cell mats.

We developed an autoradiographic screening test for hormone sensitivity of single cell suspensions of tumor tissues on cell mats, which inhibited the growth of normal cells alone. We applied this method to our newly established KSE-1 line derived from esophageal carcinoma and compared this method with well-established cytoplasmic and nuclear assays. Our assay, though taking longer to implement and providing only qualitative information, requires significantly smaller specimens than the current biochemical assay and will predict the hormone sensitivity of only viable neoplastic cells.

Leiomioma do esôfago associado à hernia de hiato. A propósito de um caso / Leiomyoma of the esophagus associated with hernia hiatal. A case report

É apresentado um caso de leiomioma esofagiano associado a hérnia de hiato e säo comentados os principais aspectos relacionados a este raro tumor, de acordo com a literatura revisada

[Dna ploidy in adenocarcinoma of Barrett's esophagus]. / Etude de l'ADN-ploïdie dans les adénocarcinomes sur endobrachyoesophage.

We have studied by flow cytometry the ADN-ploidy of 23 adenocarcinomas developed on Barrett's oesophagus operated at hospital Beaujon between 1982 and 1988. This retrospective study was done on formalin-fixed and paraffin-embedded material. Non dysplastic Barrett's mucosa was diploid in all of the 11 studied cases. Dysplastic mucosa was aneuploid in the 4 studied cases, as were the carcinomas in the same patients. Seven tumors were diploid, and 16 aneuploid. There was no relationship between the aneuploidy and the degree of tumor differentiation. Fourteen of the 15 tumors which invaded the adventitia and only 2 of the 8 tumors which were limited to the muscularis propria were aneuploid. Thirteen of 16 aneuploid and only 2 of 7 diploid tumors had lymph node invasion. Six of the 7 patients with diploid tumor were well 12 to 52 months after surgery. Eleven of the 16 patients with aneuploid tumor died, the remaining 5 were well 12 to 18 months after surgery. The ratio of aneuploid adenocarcinomas developed on Barrett's oesophagus is similar to the ratio observed in other types of solid tumors. The prognosis of adenocarcinoma in Barrett's oesophagus is poor. According to our results, the prognosis of diploid tumors seems to be better than that of aneuploid tumors. In order to determine the value of ADN-ploidy as an independent prognostic criterion, it would be of interest to study a greater number of patients with longer follow-up.

EGF and TGF-alpha, the ligands of hyperproduced EGFR in human esophageal carcinoma cells, act as autocrine growth factors.

In order to ascertain autocrine growth factors in esophageal carcinomas, we analysed expression of mRNAs and proteins for epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha) and epidermal growth factor receptor (EGFR) in 6 esophageal carcinoma cell lines. Gene alterations were also examined. All of the esophageal carcinoma cell lines expressed mRNA for EGFR and TGF-alpha genes. Interestingly, EGF mRNA of about 5.0 kb was also detected in TE-1, TE-2, and TE-8 cells. Production of protein was also confirmed by binding assay and ELISA on 3 of the 6 cell lines. The cells had a relatively high number of EGFRs and produced TGF-alpha and EGF protein at the same time. Furthermore, anti-EGF (KEM-10) and anti-TGF-alpha (WA-3) monoclonal antibodies (MAbs) inhibited spontaneous uptake of tritiated thymidine (3H-TdR) by TE-1 cells which expressed EGF, TGF-alpha and EGFR mRNA and protein. These results strongly suggest that EGF and/or TGF-alpha produced by carcinoma cells function as autocrine growth factors for human esophageal carcinomas.

DNA analysis and prognosis of digestive tract cancers.

Cytophotometric DNA analysis was performed on esophageal and gastric carcinomas. In 35 cases of mucosal and submucosal carcinoma of the esophagus, patients with types I and II (relatively regular in DNA distribution) had an uneventful postoperative course and no recurrence, whereas 3 of 15 (20%), and 5 of 9 (55.6%) with type III and type IV, respectively (widely scattered DNA distribution, died following a recurrence. Cytophotometric DNA analysis using biopsy specimens from 75 patients with esophageal cancer in various stages also showed a close relationship between the DNA distribution pattern and prognosis. However, the growth mode and the DNA ploidy of mucosal gastric cancer correlated well in the investigation of 66 cases. Thus, data of this method closely reflected the outcome in patients with digestive tract cancers. These results suggested the potential usefulness of cytophotometric DNA analysis for assessing the prognosis, even in the early stage of cancers.

Image cytometry DNA analysis of diethylnitrosamine-induced dysplasias and invasive squamous cell carcinomas of the esophagus in mice.

The nuclear DNA content was assessed by image cytometry in squamous epithelial dysplasias and invasive squamous cell carcinomas of the esophagus induced by diethylnitrosamine (DEN) in mice. The study comprises 48 lesions: 27 lesions with low grade (i.e. slight and moderate) dysplasia, 18 with high grade dysplasia (i.e. severe dysplasia and CIS), and 3 with invasive squamous cell carcinoma. In addition, 5 parallel run control specimens were also investigated. The results demonstrated that only 3.7% (or 1/27) low grade dysplasias but as much as 72.2% (or 13/18) high grade dysplasias, and all three invasive squamous cell carcinomas displayed non-diploid DNA values. Three of 18 high grade dysplasias and all three invasive squamous cell carcinomas demonstrated aneuploid cell nuclei. The results of the present work indicate that the esophageal mucosa of the mouse permit investigation--under controlled conditions--of the nuclear DNA alterations occurring during carcinogenesis in this organ. The results presented herein thus substantiate the theory of increasing DNA aberrations occurring during carcinogenesis in the human esophagus.

[A case of adenoid cystic carcinoma of the esophagus].

A surgical case of an adenoid cystic carcinoma (ACC) of the esophagus in a 75-year-old man is reported. Histologically, the tumor consisted of an ACC, a squamous cell carcinoma and a small tubular adenocarcinoma. The ACC and the tubular adenocarcinomatous regions in the submucosa and the lamina propria were continuous with the overlying squamous cell carcinoma and atypical squamous epithelium. Immunohistochemically, two types of tumor cells were detected in the ACC. One was found to have EMA-positive epithelial cells whereas the other had actin-positive myoepithelial cells. In the normal esophageal gland, actin-positive cells are found at the periphery of acini and around the layer of epithelial cells in the small duct but they have not been detected in the main duct. These findings suggest that the tumor developed from the small duct and differentiated into two directions: an ACC and a squamous cell carcinoma.

[Expression of the epidermal growth factor receptor in esophageal cancer].

The level of epidermal growth factor receptor (EGF-R) has been measured by a competitive binding assay in 45 human esophageal cancer tissues. The EGF-R level in the esophageal cancer tissue was found to be significantly higher than in specimens of gastric or colonic cancer. No correlation was observed, however between the EGF-R level and the pathological findings, such as the depth of the tumoral invasion, the degree of lymph node metastasis, and the histologic staging. Even so, the prognosis of the patients with a high level of EGF-R of more than 50 fmol/mg of protein was significantly worse than those with a low level of EGF-R and less than 50 fmol/mg of protein (p less than 0.01). Further, according to an immunohistochemical assay using anti EGF-R monoclonal antibody, EGF-R was focally stained in the basal cells and in the parabasal cells of normal esophageal mucossa, but was diffuse in almost all tumor cells of ane esophageal cancer. Thus, a good relationship with the quantitative level of EGF-R and its immunoreactivities was demonstrated, indications that EGF-R plays an important role in tumor progression and appears to be a useful prognostic factor in cases of esophageal cancer.

Myogenic expression in esophageal polypoid tumors.

Four cases of esophageal polypoid tumors composed of squamous cell carcinoma and spindle cell sarcomatous components were investigated. Squamous cell carcinoma was consistently present in the base of the polypoid lesions in all four cases and was also intermingled with spindle-shaped sarcomatous cells in two cases. Metastases in the lymph nodes were observed in two cases: one was squamous cell carcinoma with a sarcomatous component and the other consisted of a pure sarcomatous component. All tumors involved at least the muscularis mucosae. In the sarcomatous region, the tumor was composed of highly anaplastic cells with or without forming interlacing fascicles. Pleomorphism was marked and bizarre giant cell forms were frequent. Mitoses were frequently present. Immunohistochemical study revealed that the anaplastic cells in the sarcomatous component in all cases were immunoreactive to desmin, muscle actin, vimentin, and alpha 1-antichymotrypsin, but were negative for cytokeratin, even in the metastatic tumors of the lymph nodes. The immunohistochemical results favor myogenic differentiation of the anaplastic cells, and these tumors were considered to be true carcinosarcomas composed of squamous cell carcinoma and leiomyosarcoma.

[Flow cytometric analysis of DNA ploidy in esophageal and colorectal cancers--in relation to prognosis, pathologic findings, or family history of cancer].

Correlation between nuclear DNA content of tumor cells and survival, pathologic findings, as well as family history of patients was studied using paraffin-embedded materials from 85 cases of esophageal cancer and 52 cases of colorectal cancer. Nuclei were isolated from paraffin sections by the method of Hedley et al. and stained with propidium iodide. DNA index was calculated using nuclear DNA content of lymphocyte from the same patients as the external standard. Distribution of DNA indices showed a single peak in esophageal cancer, while it was bimodal in colorectal cancers. In esophageal cancers, although there was a significant correlation between survival and depth of invasion or nodal involvement, there was a very weak correlation between survival and DNA aneuploidy. Only for advanced cases, the patients who had tumors with diploidy or low ploid aneuploidy lived significantly longer than those with high ploid aneuploidy. On the other hand, in regard to colorectal cancers, the patients who had tumors with diploidy survived significantly longer than those with aneuploidy. Furthermore, patients who showed higher degree of aneuploidy tended to have poorer prognosis. There were no correlations between DNA ploidy and histologic type, depth of invasion, nodal metastasis, stage or family history of cancer.

Role of estrogen receptors in the growth of human esophageal carcinoma.

In order to investigate the role of estrogen receptors (ER) in the growth of human esophageal carcinoma, tumor tissues of ER-positive but androgen receptor (AR)-negative line of squamous cell carcinoma (ES-8) were transplanted in ten male and ten female nude mice. As control, those of a tumor line without both receptors (ES-13) derived from human esophageal cancer were used. The growth rates of transplanted tumors were estimated up to 5 weeks. The tumor growth of ES-8 line was significantly greater in males than it was in females. Such a difference was not observed for ES-13 line. Moreover, in order to investigate the effect of estrogen on the ER, tumor tissues of ES-8 line were transplanted in six oophorectomized female mice, and next, estradiol (E2) (500 micrograms/kg, 50 micrograms/kg, or none) was administered to five transplanted female mice of each group, respectively. The growth rates of tumors were enhanced in oophorectomized female mice, and significantly suppressed with the physiologic dose (50 micrograms/kg) of E2 compared to the control. The current results seem to indicate that the inhibitory effect of estrogen on the tumor line is mediated by ER.

Prognostic significance of epidermal growth factor receptor in esophageal squamous cell carcinomas.

The prognostic value of epidermal growth factor (EGF) receptor level was studied in 32 patients with esophageal squamous cell carcinoma. The EGF receptor levels of tumors were measured by iodine 125 (125I)-EGF binding assay, and the patients subsequently were divided into two groups: a group with high EGF binding capacities (greater than or equal to 2.5% of input), and a group with low EGF binding capacities (less than 2.5% of input). The cumulative survival rates for the two groups were calculated by the Kaplan-Meier method. The generalized Wilcoxon test indicated that the survival rate of the high EGF binding group was significantly lower than that of the low EGF binding group (P less than 0.05). In tumors from two patients with the highest EGF receptor levels, EGF receptor gene amplification was observed. These patients developed mediastinal lymph node metastasis and died 4 and 11 months after surgery, respectively. These results suggest that elevated EGF receptor level is a significant prognostic indicator for esophageal squamous cell carcinoma.