Hiromichi Sumiyoshi

Expression of epidermal growth factor in human tissues. Immunohistochemical and biochemical analysis.

The expression of epidermal growth factor (EGF) was examined on various human tissues by radioimmunoassay, immunohistochemistry and Northern blot analysis. Immunoreactive EGF was found in most of the human tissues by radioimmunoassay at various levels. Large quantities of EGF were detected in the kidney and thyroid gland. Immunohistochemically, EGF immunoreactivity was detected mainly in the epithelial cells of the lung, stomach, duodenum, pancreas, kidney, pituitary gland, thyroid gland, mammary gland, ovary, uterus and placenta. Weakly EGF-positive cells were also found in the adrenal gland. The results of EGF-immunostaining were not always consistent with the data from radioimmunoassay. We consider that the amount of EGF measured by radioimmunoassay reflects the density of EGF-positive cells in the tissues and the concentration of EGF in individual EGF-positive cells. Furthermore, EGF mRNA was expressed in the salivary gland, thyroid gland, mammary gland and kidney. It is thus evident that EGF is produced by a variety of human tissues. The kidney expressed exceptionally high levels of EGF mRNA which was about one-tenth of the expression in mouse submandibular gland, suggesting that most of EGF in the urine is produced and secreted by the epithelial cells of renal tubules.

DISTRIBUTION OF COLLAGEN TYPES I, III, AND V IN FIBROTIC AND NEOPLASTIC HUMAN LIVER

The distribution of collagen types I, III, and V in normal and fibrotic human livers and hepatocellular carcinoma was studied by indirect immunofluorescence procedure using type specific antibodies. Type I collagen as well as type III collagen was present in normal liver within the portal tracts and along the perisinusoidal spaces. Basement membrane collagen, type V collagen, was demonstrated only around the bile ducts and vessels of the portal tracts and central veins. In fibrotic liver, both type I and III collagens were found in increased amounts in fibrotic areas. In fibrous septa of active cirrhosis, however, type I collagen as well as type III collagen was abundant, whereas in inactive cirrhosis type I fibers were predominant. Type V collagen was observed in the walls of proliferative bile ductules and vessels in the fibrotic liver, and also along the sinusoids in the periportal areas. In hepatocellular carcinoma, each type of collagen was distributed regularly along the sinusoid‐like vascular channels within the tumor.

Distribution of collagen types I and III and basal lamina in human gastric carcinoma: an immunohistochemical and electron microscopic study

Collagen types I and III were examined immunohistochemically in 32 cases of gastric carcinoma classified as poorly differentiated adenocarcinoma with scirrhous stroma, well differentiated adenocarcinoma with intermediate stroma, or poorly differentiated adenocarcinoma with medullary stroma. In the stroma of scirrhous carcinoma, types I and III collagens were distributed abundantly in fibrillar or granular patterns with little difference in the intensity of staining. In well differentiated adenocarcinoma, type I collagen was diffusely distributed in the stroma with type III collagen distributed sparsely. In poorly differentiated adenocarcinoma with medullary stroma, the two types of collagen were only found around capillaries, constituting the tumor interstitium. Electron microscopic examination of scirrhous carcinoma showed tumor cells partially covered with fibroblasts, and discontinuous basal lamina, collagen fibers and microfibrils present between tumor cells and fibroblasts. In well differentiated carcinoma, tumor cells were surrounded by fibroblasts, and well developed basal lamina was observed beneath the tumor cells. In poorly differentiated carcinoma with medullary stroma, the stroma consisted of capillaries and very few fibroblasts with discontinuous basal lamina occasionally being present between tumor cells and fibroblasts.

Argyrophil cells in early gastric carcinoma: an immunohistochemical and ultrastructural study.

SummaryEighteen argyrophil cell carcinomas in 101 early gastric carcinomas were examined histologically, ultrastructurally, and immunohistochemically for polypeptides, carcinoembryonic antigen (CEA), lysozyme, and human chorionic gonadotrophin (hCG). Seven of these 18 tumors had gastrin, and two of seven tumors also contained somatostatin. In all of these 18 tumors CEA were demonstrated. Seven had lysozyme and five of seven tumors also contained gastrin; hCG were present in four of 18 tumors and two of four tumors had gastrin, CEA, mucin, and lysozyme simultaneously. Argentaffin cells were found in seven of 18 tumors. Of the above seven tumors containing gastrin, three had argentaffin cells. Ultrastructurally, several types of secretory granules were noted and tumor cells resembling D1-or P cells were present in nine of the 18 tumors. Macroscopically, many of the tumors showed IIc or IIc+III type. Histologically, the 18 tumors consisted of six well differentiated adenocarcinomas and 12 poorly differentiated adenocarcinomas including signet-ring cell carcinoma. These 12 tumors frequently developed in the stomach of young females. In view of our previous investigations, it was suggested that the IIc-type argyrophil cell carcinoma histologically showing poorly differentiated adenocarcinoma may be related to scirrhous carcinoma of the stomach.Zusammenfassung18 Argyrophilzellencarcinome aus insgesamt 101 Frühcarcinomen des Magens wurden lichtmikroskopisch, elektronenmikroskopisch sowie immunhistochemisch mit Antiseren gegen Polypeptide, CEA, Lysozyme und hCG untersucht. 7 dieser Tumoren enthielten Gastrin und 2 unter ihnen außerdem Somatostatin. In allen 18 Tumoren wurde CEA nachgewiesen, 7 von diesen 18 Tumoren zeigten Lysozyme, darunter 5 auch noch Gastrin. In 4 Tumoren wurde hCG beobachtet, und in 2 aus dieser Reihe von 4 fand sich gleichzeitig Gastrin, CEA, Mucin sowie Lysozyme. Argentaffine Zellen wurden in 7 von 18 Tumoren beobachtet. Drei von 7 Gastrin enthaltende Tumoren hatten mehr oder weniger argentaffine Zellen. Elektronmikroskopisch wurden unterschiedliche Sekretgranula beobachtet und 9 von 18 Tumoren waren D1 oder P Zellen ähnlich. Makroskopisch entsprach die Mehrzahl der Tumoren dem IIc oder IIc+III Typ. Histologisch waren von diesen 18 Tumoren 6 gut differenzierte und 12 wenig differenzierte Adenocarcinome, einschließlich Siegelring-zellencarcinomen. Sie waren öfters im Fundusbereich jüngerer Frauen lokalisiert. Unter Heranziehung unserer schon publizierten Untersuchungsbefunde wird vermutet, daß IIc-Typ Argyrophilzellencarcinome mit dem histologischen Bild des wenig differenzierten Adenocarcinoms dem scirrhösen Carcinom zugeordnet werden kann.

Gut endocrine cells in rat stomach carcinoma induced by N-methyl-N'-nitro-N-nitrosoguanidine.

SummaryGut endocrine cells in a total of 18 gastric adenocarcinomas in inbred Wistar rats induced by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and gastrin or serotonin, were examined histologically, ultrastructurally, and immunohistochemically for gastrin, somatostatin, calcitonin, glicentin, and serotonin. A large number of argyrophil cells were observed in 17 tumors (94.4%) and 14 tumors (77.8%) had argentaffin cells. Immunohistochemically, C-terminal fragment of gastrin (G17) immunoreactivity was observed in 15 (82.2%) out of the 18 tumors, but 3 G17-positive tumors had no G 34 immunoreactive cells in rats treated with MNNG plus gastrin. Serotonin immunoreactivity was detected in 14 tumors (77.8%). Somatostatin immunoreactivity was detected in 7 of the 11 tumors (63.6%) in rats treated with MNNG plus gastrin whereas no tumor in rats treated with MNNG plus serotonin had somatostatin, the difference of the incidence being significant (P<0.05). One endocrine cell carcinoma which consisted mainly of serotonin-producing cells was observed in a rat treated with MNNG plus serotonin. Calcitonin and glicentin immunoreactivity was not demonstrated in any tumors. Ultrastructurally, three types of endocrine granule were found in the tumor cells.These data suggest that hormonal environment in stomach carcinogenesis may influence the expression of endocrine cells within the tumors.

EXPRESSION OF Ha-ras ONCOGENE PRODUCT IN RAT GASTROINTESTINAL CARCINOMAS INDUCED BY CHEMICAL CARCINOGENS

The expression of Haras oncogene product in rat gastrointestinal carcinomas induced by N‐methyl‐N‐nitro‐N‐nitrosoguanidine (MNNG) or 1, 2‐dimethylhydrazine (DMH) was studied by Western blotting and immunohistochemistry using anti‐Haras p 21 oncoprotein antibody. In Western blotting, high levels of c‐Haras p 21 were found in serially transplantable rat duodenal carcinomas induced by MNNG and rat colon carcinomas induced by DMH. mmunohistochemically, c‐Haras p21 immunoreactivity was detected in 3 (16.7%) of 17 MNNG‐induced stomach carcinomas and in 21 (63.6%) of 33 DMH‐induced colon carcinomas, respectively. In the colon carcinomas, c‐Ha‐ras p 21 immunoreactivity in deeply invasive tumors was stronger than that in superficially invasive tumors and was expressed in all subserosal tumors. Moreover, all of the metastatic colon carcinomas had c‐Ha‐ ras p 21 immuno‐reactive tumor cells. These findings suggest that c‐Ha‐ ras p21 expression plays an important role in tumor proliferation, invasion and metastasis of DMH‐induced colon carcinoma. ACTA PATHOL. JPN. 37: 1731–1741, 1987.

Effects of a Protein-Free Diet on the Rat Stomach

The effects of a protein-free diet containing a mixture of free amino acids on the stomach were studied using male Wistar strain rats. In the forestomach, thickening of mucosa and hyperkeratosis appeared and papillomas frequently developed, but they did not occur in the control rats fed a semipurified diet. Mucous neck cells increased and parietal cells and chief cells decreased in the fundus. Goblet cells occurred in the antral glands, and intestinal metaplastic glands developed in the late stage of the experiment. The number of deoxyribonucleic acid-synthesizing cells in the glandular stomach was significantly higher than that of the control. Moreover, in the antrum, somatostatin-producing cells increased in the early stage and gastrin-producing cells decreased in the late stage. These results suggest that dietary protein might play a crucial role in the differentiation of gastric epithelium and the development of intestinal metaplasia in the stomach.