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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.

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.

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.

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.

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'.

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.

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.

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.

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.

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.

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.

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.

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.

Non-haem iron concentrations were measured in 65 specimens of liver obtained from 50 male and 15 female black adults who died from carcinoma of the oesophagus in the Johannesburg area between 1971 and 1980. The results were compared with those obtained in two earlier studies on subjects who died from other causes. The first was carried out on 220 men and 164 women in 1959/1960 at Baragwanath Hospital, Johannesburg, and the second series, which consisted of 248 men and 181 women, was obtained at the same hospital in 1976. During the intervening period there was a marked drop in the incidence and severity of iron overload in urban blacks. A direct comparison between subjects with carcinoma of the oesophagus and the 1976 group showed that the geometric mean hepatic iron concentrations in the carcinoma patients were significantly higher in the 40-49, 50-59 and over-60 age groups. While geometric mean hepatic iron concentrations in the various age groups were also higher than those obtained in the 1959/1960 study, the differences only reached statistical significance in subjects over the age of 60 years. Seven of the 15 women had significant hepatic iron deposits (greater than 0,5% dry weight). These results suggest that the excessive consumption of home-brewed alcoholic drinks contaminated with iron may be directly or indirectly associated with the development of carcinoma of the oesophagus in urban black adults.

Cytofluorometric analysis of nuclear DNA content was performed in carcinoma cells of the stomach and the esophagus and the exclusive four types of the DNA histogram patterns were established. In 33 patients with carcinoma of the stomach 7 (87.5%) out of 8 with Type I and 3 (50%) out of 6 with Type II in the DNA histogram pattern were the superficial types of carcinomas. Eighteen (81.8%) out of 22 patients with advanced gastric carcinoma belonged to Type III or IV in the DNA histogram pattern and there was only one of the superficial spreading carcinoma seen in these Types. The superficial spreading carcinoma (Stout 10) is generally reported to take more favorable clinical course than the usual advanced ones and the above data were supported that the DNA histogram pattern was associated with prognostic important property of invasive growth. In 34 patients with carcinoma of the esophagus 4 out of 7 with Type I in the DNA histogram pattern have been living more than 3 years following surgery with poor lymphatic permeation (14.3%) around the tumor. While, 6 out of 9 with Type IV had an early recurrence of cancer causing postoperative death within a year after surgery with high incidence of lymphatic permeation (77.8%) around the tumor. Thus, the DNA histogram pattern in the cytofluorometric DNA analysis of the tumor cells seems to give valuable information about the malignancy degree of the individual tumor, which is not possible to obtain only from the cytomorphological features.

The Feulgen-DNA content of squamous carcinoma cell nuclei from the human esophagus was assessed in punch biopsies from 47 untreated patients. Forty-four of the 47 biopsies (93.6%) demonstrated aneuploid cell populations, and the remaining 3 (6.4%) were non-diploid. Previous studies have demonstrated that in esophageal dysplasias adjacent to invasive squamous cell carcinoma, DNA in single cells is substantially altered. Thus the process of esophageal carcinogenesis can be monitored not only by histological changes, but also by DNA aberrations in single cells. Quantitative DNA measurement appears, therefore, to be a complement to the histological evaluation of esophageal lesions with suspected, but not unequivocal, evidence of neoplastic growth.

The role of epidermal growth factor (EGF) and its receptors in human cancers was studied using 24 human cell cultures including 15 of squamous cell carcinoma (SCC) of the skin, oral cavity, and esophagus. EGF was found to inhibit the growth and colony formation of all the SCC cells at doses that are mitogenic in many other cells, including epidermal keratinocytes and dermal fibroblasts. This inhibitory effect of EGF on SCCs was specific, because EGF did not inhibit and in some cases slightly stimulated the growth of other tumor cells, such as adenocarcinomas of the stomach, cervix, and breast and sarcomas. The amounts of EGF receptors on these SCC cells were measured by immunoprecipitation of labeled proteins with anti-EGF receptor polyclonal antibody and binding assay of membrane preparations using 125I-EGF. Of 13 SCC cell cultures tested, all except 3 of esophageal SCC showed higher levels of EGF receptor than normal epidermal keratinocytes, which contain 1.5 X 10(5) binding sites/cell. In general, SCCs of the skin and oral cavity had large amounts of EGF receptor on the order of 10(6)/cell, whereas the receptor of esophageal SCCs was on the order of 10(5)/cell. Some SCC cells had about twice as many EGF receptors as A431 cells. The values for the equilibrium dissociation constant (Kd) of these cells were on the order of nM. The sensitivity to the inhibitory effect of EGF correlated well with the elevated level of EGF receptors in 12 SCC cell lines, and higher significance was obtained when data on esophageal SCCs were excluded. The present observations suggest that EGF and EGF receptors play a role in the development of SCCs.

Although it is widely known that early esophageal cancer (n-) patients have a favorable prognosis after surgery and the superficial esophageal cancer (n+) patients have a poor one, the differences between them have not been found out under the microscopic examination of the primary lesion. Cytophotometric DNA analysis of the primary lesion was done to search for the differences between early and superficial esophageal cancer. Significant differences were obtained in the mean DNA value (p less than 0.01), the percentage of nuclei beyond tetraploid (p less than 0.001) and the product of 2 indices above (p less than 0.001). The result suggests that cytophotometric DNA analysis is a useful means for differentiating the early esophageal cancer from the superficial esophageal cancer and that the most suitable perioperative combined therapy can be chosen through the Feulgen-stained biopsy specimen.

The purpose of this paper is to assess the clinical and histopathological significance of tumors at the esophago-gastric junction (EGJ). The biological behaviour of these tumors was studied, with regard to the infiltrative pattern and mitotic index, in 15 patients with tumors at the EGJ. Twenty-seven tumors at the cardia, with or without esophageal invasion, were chosen for DNA analysis at their most proximal and deepest apexes. The esophageal and gastric apexes of the tumors had the same infiltrative pattern in 14 out of 15 cases and this tendency was not altered with further stratification of the anatomical layers. In comparing the deviations of the stem lines, the relative frequency of nuclei with DNA over 4n and 6n and aneuploid from the DNA histograms between the esophageal and the deepest apexes of the tumor revealed no statistically significant differences, depending on esophageal invasion. From these results, the specificities of EGJ toward tumor invasion or activity were not regarded as consequential, and the EGJ couldn't be regarded as a barrier against tumor invasion.

A case of primary adenoid cystic carcinoma of the esophagus was reported. A 54-year-old male patient had a tumor in the middle third of the esophagus. Barium swallow and endoscopic examination showed a protruding tumor with a shallow ulceration in its vertex. Histological examination of the surgically removed material revealed that it was largely confined to the submucosal layer, with no metastasis to lymph nodes. The tumor cells presented three distinct patterns: globular nest with irregular cystic spaces, trabecular nest, and true tubule. Electron microscopic study revealed no microvilli or triad along cystic spaces in the globular nests. Histochemical study disclosed that cystic spaces and interstitium were rich in glycosaminoglycans. The luminal surface of the true tubules of the tumor, on the other hand, was characteristically lined by sialomucins. The trabecular type-nests differed from the globular nests in showing GSA-II reactivity and containing abundant retinol-binding protein. These results indicate that the three patterns represented different differentiations of the tumor cells. Twenty-three cases of esophageal adenoid cystic carcinoma were found in the literature from 1950 to 1983 and discussed collectively.

The mucin profile of 24 endoscopic biopsies of heterotopic gastric mucosa (HGM) of the upper esophagus in adults and a control group of ten cases of Meckel's diverticula containing heterotopic gastric mucosa were studied with two combined histochemical methods: alcian blue pH 2.5/PAS and high iron diamine/alcian blue pH 2.5. The clinical and light microscopic features of the 24 HGM cases were also reviewed. In addition to overall secretion of neutral mucins by the 24 HGM cases, mucin histochemistry showed prominent secretion of acidic mucins in 19 of 24 HGM cases (79%), with sulphomucins in 11 of 24 HGM cases (45.8%). This mucin profile of HGM was unlike that of either normal gastric mucosa or heterotopic gastric mucosa in Meckel's diverticula. Moreover, a comparison between the mucin profile and clinical features of HGM and Barrett's esophagus showed certain similarities. The data suggest a physiopathologic link between HGM and Barrett's esophagus.

The nuclear DNA content was microspectrophotometrically measured in 16 resected esophagi having dysplasia, carcinoma in situ and/or early invasive squamous carcinoma. First, the epithelial thickness in (1) normal squamous esophageal epithelium and (2) dysplasia-carcinoma in situ areas was divided into three equal compartments (i.e., basal-parabasal, intermediate and superficial) in five cases. In the normal epithelium, while some of the nuclei in basal-parabasal normal squamous cells had elevated DNA values (corresponding to the natural DNA replication in these cells), intermediate and superficial (nonreplicating) normal squamous cells showed a more definite clustering about the 2c value. In the nonnormal epithelium, the percentage of cells with DNA levels exceeding the normal tetraploid value was highest for the intermediate zone. Therefore, in all 16 cases, normal intermediate cells were measured as internal controls, against which the DNA levels of cells in the intermediate compartment in the areas of dysplasia and/or carcinoma in situ were compared. In areas of dysplasia, two different DNA patterns were observed: one clustering around the normal diploid region and the other with aneuploid values. While the former corresponded to some of the lesions considered by conventional histologic examination to be slight and moderate dysplasias, the aneuploid pattern corresponded to the remaining slight and moderate dysplasias as well as to the severe dysplasias. The possibility that "diploid dysplasias" are reactive (i.e., nonneoplastic) lesions due to chronic inflammation or are "dormant" nonprogressive dysplasias, while aneuploid dysplasias are more aggressive lesions, seems to be substantiated by the fact that all areas with carcinoma in situ or with microinvasive squamous carcinoma had aneuploid nuclei.

Millet bran is a component of the diet in the area of highest oesophageal cancer incidence in northern China. Millet bran was found to contain up to 20% by weight of silica; some of this silica occurs as friable sheets or sharply-pointed fibres. These types of silica in millet bran are the most likely source of an unusual contamination with fragments of silica found in the oesophageal mucosa surrounding tumours in patients in northern China. A group of mucosal samples analysed together contained over 5,000 particles/g (100 parts per million by weight), ten times as many as were found in tissue from normal controls taken at necropsy in London. The modal diameter was 10 microgram (1-70 microgram). The particles were in the body of the mucosa and were not simply a surface contaminant. Silica fragments and fibres of similar size originating from other plant species occur in the diet in the two other regions of greatest incidence of oesophageal cancer, the Transkei and Iran. If such fragments enter the mucosa, they must cause some degree of trauma, and they may also be able to stimulate proliferation by providing anchorage. These findings suggest the possibility that silica particles might be involved in the aetiology of oesophageal cancer.

The 'epidemiological necropsy' is a newly proposed research strategy in which the size and composition of the epidemiological reservoir of undetected disease is estimated from the relative frequency of necropsy surprise patients, in whom the disease was not suspected during life. The current study was done to help validate a basic premise of the strategy. We examined the surprise necropsy discovery of two upper gastrointestinal malignancies: oesophageal cancer, for which an undetected reservoir would not be expected because the cancer has little room to grow, and gastric cancer, for which a sizeable reservoir might be anticipated. In a review of 15,812 necropsies during 1953-1982 at Yale-New Haven Hospital, 70 cases of oesophageal cancer were identified postmortem. Except for five surprise cases, located at the gastro-oesophageal junction where there is room to grow, no oesophageal cancer reservoir was found during the 30-year period. In the same secular period, however, a distinctive set of reservoir cases was found among 162 necropsy instances of gastric cancer. About two-thirds of these gastric cancers had been previously diagnosed during life, but the remainder consisted of either necropsy surprise cases or patients with wrong primary cancer diagnoses during life. The secular rates of occurrence remained stable and similar in both sexes for surprise gastric cancer cases during the three 10-year periods from 1953-1982, but no wrong primary diagnoses occurred during 1978-1982, after the introduction of improved methods of premortem diagnosis. The results help validate the cancer reservoir theory for malignancies that have room to grow, and confirm the concept that the 'epidemiological necropsy' can reflect qualitative and quantitative changes in cancer reservoirs.

The correlation of 5-year survival rate with various clinical and histopathological factors was studied using univariate and multiple analyses of 128 patients who had undergone resection for esophageal carcinoma between 1965 and 1978 in the Department of Surgery, Kyushu University Hospital. The depth of penetration, lymph node metastasis, lymphatic or vascular invasion, and INF had a significant correlation with 5-year survival in the univariate analysis; however, only depth of penetration and lymph node metastasis were prognostic factors with a significant difference, in the multivariate analysis. In 55 patients in whom the cell nuclear DNA content had been determined, the DNA pattern was the greatest prognostic factor (p less than 0.01), in multivariate analysis. We propose that the DNA distribution in the malignant cells should be examined as a most pertinent prognostic factor.

Polypoid carcinoma of the esophagus is a rare variant of squamous carcinoma, which occurs typically in elderly patients who present with symptoms of dysphagia. This tumor has been given a variety of names, including spindle cell carcinoma, carcinosarcoma, and pseudosarcoma. These designations reflect the controversy surrounding the nature of the spindle cell component of this lesion. We have studied a case of polypoid carcinoma of the esophagus with immunoperoxidase staining for keratin which has been shown to be specific for epithelial cells. Positive staining for keratin was demonstrated in both the carcinomatous and spindle cell "sarcomatous" appearing components of the tumor, supporting the contention that this tumor is of entirely epithelial origin.

Intraperitoneal di(2-hydroxypropyl)-Nitrosamine (DHPN) caused a high incidence of oesophageal squamous carcinoma in male Wistar rats, particularly in rats killed 11 or more months after the start of injections. No control rats (injected intraperitoneally with saline) developed an oesophageal neoplasms. Histopathologically, the tumours were moderately well differentiated. Histochemical studies showed minor increases in mucin staining and mast cell population and a marked increase in bacteria in tumour-bearing oesophaguses. Electron microscopy showed the tumours to be similar to, but to differ in some respects from squamous carcinomas at other sites in humans. The possible implications of this work for human disease are twofold. It could provide a model for further study of aspects of oesophageal carcinoma and it serves to remind us that all potential oesophageal carcinogens need not act during swallowing.

The 8-nm keratin filament is a major component of the cytoskeleton of epithelial cells and epithelial-derived cancers (carcinomas). Recently, it has been shown that the pattern of keratins produced by an esophageal epithelial cell undergoes change upon malignant transformation. In order to evaluate the potential importance of these differences in providing improved diagnostic techniques for pathology, we have investigated the consistency of the patterns of keratins expressed in normal esophageal epithelium, squamous cell carcinoma (SQCC) of the esophagus, and cultured esophageal epithelial cells. In six patients, the keratin pattern expressed by SQCC of the esophagus and corresponding normal esophageal epithelium was consistently different as judged by immunoblot analysis of electrophoretically separated protein extracts. Whereas the SQCCs typically expressed major keratins with molecular weights of 58,000, 56,000, 50,000, and 46,000, the normal esophageal epithelium produced two major keratins with molecular weights of 58,000 and 52,000 and a minor keratin with a molecular weight of 56,000. When normal esophageal epithelial cells were grown in tissue culture, their keratin pattern changed, and keratins with molecular weights of 58,000, 56,000, 52,000, 50,000, 46,000, and 40,000 were expressed. Although some minor variations in keratin patterns were seen, the major differences in keratin pattern expressed by normal esophageal epithelial tissue, SQCC of the esophagus, and cultured esophageal cells were consistent and reproducible.

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.

In a series of 79 cases of primary esophageal carcinoma resected at The Center for Adult Diseases, Osaka, there were six tumors with specific histopathologic features valid for the diagnosis of argyrophil cell carcinoma. Of the 6 tumors, 3 were studied electron microscopically and assay for ACTH content was performed on 4 tumors. Clinically, the ages of the 6 patients ranged from 56 to 71 years; two were women and four men. Four of the 6 patients died with widespread tumor recurrences within 9 months of operation. Microscopically, the 6 tumors were composed largely or almost entirely of small, spindle-shaped cells resembling those of oat cell carcinoma of the lung, and were characterized by the arrangement of tumor cells in solid sheets or anastomosing cords, the presence of argyrophil tumor cells, and the deposits of amyloid. Electron microscopically, the three tumors contained neurosecretory-type granules. Using bioassay or radioimmunoassay ACTH activity in the tumor tissues was detected in 3 out of the 4 tumors determined. From the light and electron microscopic characteristics and the assay evidence, it seems reasonable to conclude that the 6 tumors are endocrine polypeptide producing tumors (apudomas) that arise from argyrophil cells normally found among the basal cells of the esophageal mucosa, and that they represent a distinct histopathologic entity clearly distinguishable from other types of esophageal carcinomas.

A series of esophageal lesions, mild, moderate and severe dysplasia, carcinoma in situ (CIS), early and advanced squamous cell carcinoma were induced in rats with N-methyl-N-amylnitrosamine (MAN). To evaluate the biological nature of each lesion, the ploidy level was estimated by microspectrophotometrical measurement of cell nuclear DNA content. DNA distribution patterns were classified into types I, II, III and IV, according to the degree of dispersion and the peak modal value on the DNA histogram. The incidences of type III of high ploidy in early cancer, CIS and severe dysplasia were 3/11 (27.3%), 5/13 (33.3%) and 4/16 (25%), respectively. On the other hand, in moderate and mild dysplasia, 15/16 (93.8%) and 20/21 (95.2%) were low ploidy (types I and II), respectively. The mean DNA content of advanced and early cancer, CIS and severe dysplasia were 3.88c, 3.34c, 3.24c and 3.13c, respectively, while those of moderate and mild dysplasia were near diploid, showing 2.67c and 2.51c, respectively. These findings indicate that the biological nature of severe dysplasia may be considered as serious a lesion as cancer, in terms of DNA analysis. Cytophotometric DNA analysis aids the evaluation of various degrees of dysplasia and carcinoma of the esophagus.

The concentration of cellular retinoic acid binding proteins (CRABP) was determined in the cytosol of normal esophageal tissue and in esophageal carcinomas. Unlike the reported results for human breast, colon, melanoma, or oropharynx cancers, the CRABP levels in esophageal cancers were either undetectable or contained levels of CRABP which were significantly lower than that of adjacent histologically disease-free tissue (P less than 0.005). Moreover, there was no difference between the normal mucosa of cancer or noncancer patients with regards to the CRABP concentration. The absence of CRABP in the cancer tissue was not dependent on the degree of differentiation. These results indicate that the CRABP disappears when the normal mucosa becomes malignant. If such a change is also demonstrated in known premalignant conditions of the esophagus, CRABP could serve as a diagnostic biochemical marker for early detection of this cancer.

Fifty five formalin-fixed, paraffin-embedded, pronase-pretreated esophageal carcinomas and their metastatic nodes were studied immunohistochemically with peroxidase-antiperoxidase technique for keratins using polyclonal antibody (DAKO), antibody against keratins of keratinocyte (KL1), against high molecular weight-keratin (EY904), and against low molecular weight-keratins (EY902, PKK1). The suprabasal layer of the esophageal epithelium was stained diffusely with DAKO, KL1 and EY902 and faintly or locally with EY904 and PKK1. Esophageal ducts were positive with any antibodies. We classified staining pattern of the esophageal carcinoma according to the localization and proportion of the positive cells in the cancer nests. No significant correlation of the staining pattern was found between the primary lesions and metastatic nodes. The proportion of the EY904-positive cells correlated significantly with the degree of lymph node metastasis, histologic stage, and prognosis by rank correlation method and generalized Wilcoxon test, but not with the depth of invasion nor histologic degree of differentiation. The proportion of DAKO-positive cells correlated significantly only with histologic degree of differentiation. It was supposed that esophageal carcinomas with high molecular weight-keratin had less tendency to metastasize to nodes.

Biopsy tissues from a 68-year-old Japanese man with metastases to axillary lymph nodes of a recurrent esophageal carcinoma were adapted to cell culture conditions and a continuously growing tumor cell line was developed. Immunohistochemical staining revealed that these cells contained keratinous material and the electron microscopic study revealed the presence of tonofilaments. Thus, this line, designated the KSE-1 line, was considered to have originated from metastatic squamous cell carcinoma of the esophagus. This line has a binding content of 4.2 fmol/mg protein for the estrogen receptor and 2.2 fmol/mg protein for the testosterone receptor. By measurement of cell number and thymidine incorporation, the growth rate of this line was found to be moderately responsive to these hormones, being inhibited by estrogen and enhanced by testosterone at concentration levels between 10(-13) and 10(-8) and 10(-13) and 10(-6) mg/ml, respectively.

Trace element concentrations were determined in surgically resected, cancerous human oesophageal tissue by proton-induced X-ray emission (PIXE) analysis. Each sample was divided into six zones, E3 and E4 formed the central tumour region, E1 and E2 were proximal to the oesophagus and E5 and E6 distal to the oesophagus. Malignancy was confirmed histopathologically in all cases. Samples were dissolved in NaOH solution and pipetted onto a thin mylar film for analysis by a proton beam from a Van de Graaff accelerator. Titanium, Fe, Cu, Zn, As, Rb, Br and Pb were detected in varying concentrations. Of these, Ti, Fe, Cu, Zn and As showed specific behaviour in the six zones.

A total of 353 samples of gastric juice was collected from Lin-xian subjects who were examined by endoscopy. NDMA, NDEA, NMBzA, Npyr, Npip, NSAR and other unknown compounds were detected in the fasting gastric juice. NMBzA, Npyr, Npip and NSAR can induce esophageal cancer in animals. Among the concentrations of N-Nitrosamines in gastric juice, the level of NDMA was the highest, its mean value 17.09 ppb; the level of NDEA stood next with a mean value of 6.95 ppb; the amounts of NMBzA, Npyr and Npip were 4.77, 2.45 and 1.30 ppb, respectively. A positive correlation was found between the amount of N-Nitrosamines in gastric juice and the various lesions in the esophageal epithelium of the subjects, the amount of N-Nitrosamines in gastric juice from subjects with normal esophageal epithelium was lower than that from subjects with marked dysplasia or carcinoma of esophagus. This finding lends further support to their possible involvement in esophageal cancer development.

DNA distribution of 4 early esophageal cancers and their epithelial adjacent to cancer was analysed using photocytometry technique. In the tissues around the cancer, DNA content of the normal squamous epithelium always showed diploid (2 c) or proliferating diploid (less than 5 c). DNA content of 3 cases of the dysplastic cells in three specimens was predominantly proliferating diploid, comprising 68%, 38.2% and 59.4% but the aneuploid (greater than 5 c), 8%, 2.9% and 21.8%, respectively. In 4 cases of carcinoma in situ, the aneuploid increased to 86.7%, 68.7%, 17.2% and 28.6%, markedly higher than the dysplastic cells. In 3 cases of early esophageal cancer, the aneuploid accounted for 58.8%, 51.1% and 13.6%, similar to carcinoma in situ. The results indicate that in the dysplastic cells, there is a relative high ratio of the aneuploid cells except the increased proliferating diploid. This phenomenon could explain the unstability for dysplasia and might be the earliest manifestation of malignant change of the epithelial cells.

In order to grade objectively and characterize dysplastic and precancerous esophageal epithelium its DNA content was measured by cytofluormetric methods and compared to normal and cancerous esophageal epithelium. This yielded the following results. With transition of the esophageal eipthelium from mild dysplasia to severe dysplasia and finally to in situ carcinoma, Feulgen-DNA values showed patterns characteristic of a tetraploid population. They lacked prominent peaks which were usually observed with invasive carcinomas. Dominant near-tetraploid population and definite tetraploid-octoploid populations were characteristic of severe dysplasia or carcinoma. The mean Feulgen-DNA values were significantly larger in severe dysplasia than in the lesser grade of dysplasia as well as the normal epithelium. However, this was not the rule in the full blown carcinomas. It would appear that the esophageal cytophotometric patterns are analogous to those previously observed in the skin and uterine cervix.

Eight cases of esophageal carcinoma with prominent spindle cells (carcinosarcoma or pseudosarcoma) were studied using the avidin-biotin immunoperoxidase method and monoclonal antibodies to various keratins and vimentin. In all eight cases positive immunoreactivity for keratin was found in carcinomatous areas and for vimentin, in the spindle cells. It is interesting that five cases demonstrated focal immunoreactivity to keratin in the spindle cell component. Trace positivity to vimentin was seen in the carcinomatous areas in one case. These findings are consistent with the hypothesis that esophageal carcinoma with prominent spindle cells is of epithelial origin and may represent a morphologic variant of squamous cell carcinoma.

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.

Clinically it is well known that the incidence of esophageal cancer is much higher in men that in women. Further, the prognosis of the esophageal cancer is worse for men than for women. This seems to be dependent on the difference of environment of the sex steroid. In this paper, the androgen receptor (AR) has been measured in 21 cases of human esophageal cancer. Two cases of esophageal cancer that were implanted into nude mice were AR positive. In this EH-1 cell line, the binding sites were increased and the tumor growth was enhanced by testosterone administration, but the tumor growth was not inhibited by estrogen administration.