Setsuko Hatakeyama

Expression of bone morphogenetic proteins in colon carcinoma with heterotopic ossification.

Here we report the case of a 50‐year‐old woman with adenocarcinoma of the colon, showing heterotopic ossification. The patient was referred to our hospital for investigation of anemia secondary to occult gastrointestinal blood loss. By colonoscopy, an irregular polypoid mass was found in the ascending colon. A biopsy of the lesion revealed moderately to poorly differentiated adenocarcinoma with heterotopic ossification. A right hemicolectomy was done and revealed areas of heterotopic bone within the tumor, but no ossification was evident in the metastatic lesions within the mesenteric lymph nodes. The formation of heterotopic bone in gastrointestinal tumors is rare and its exact mechanism is unknown. Immunohistochemical localization of bone morphogenetic proteins (BMP), known to be primary inducers of new bone formation, was determined. BMP‐5 and ‐6 were prominent in the cytoplasm of tumor cells, and they stained weakly in osteoblast‐like cells adjacent to newly formed bone. Cytoplasmic staining for BMP‐2 and ‐4 was weak in tumor cells, osteoblast‐like cells, and stromal fibroblast cells. BMP may play an important role in heterotopic ossification in colon adenocarcinoma.

Expression of bone morphogenetic proteins of human neoplastic epithelial cells

Bone morphogenetic proteins (BMPs) are crucial factors of osteogenesis. We investigated the expressions of BMP subtypes in human salivary adenocarcinoma cell line (HSG‐S8), tongue squamous cell (HSC‐4) and gingival squamous cell (Ca9‐22) carcinoma cell lines, gastric poorly differentiated adenocacinoma cell (MNK45) and signet ring cell (KATOIII) carcinoma cell lines, rectal adenocarcinoma (RCM‐1, RCM‐2, and RCM‐3), and thyroid (8505C) and bladder (T24) carcinoma cell lines by reverse transcription‐polymerase chain reaction (RT‐PCR). RT‐PCR disclosed that BMP‐1 was expressed in all cell lines examined, and BMP‐2 was amplified in almost all cells except MKN45. Two squamous cell carcinomas, HSC‐4 and Ca9‐22, and KATOIII expressed only BMP‐1 and BMP‐2. MKN45 did not express BMP‐2, but expressed BMP‐7 and weakly BMP‐4 and BMP‐5. In addition to the expression of BMP‐1 and BMP‐2, three rectal adenocarcinoma cell lines commonly expressed BMP‐7, and HSG‐S8 expressed BMP‐6. These findings indicated that the neoplastic epithelial cells possessed a rather great potency to express BMP mRNAs. On the other hand, among these carcinoma cells, HSG‐S8 solely induced bone in nude mouse tumors, and HSC‐4 and KATOIII contained many calcified masses in tumors while the rest did not induce either.

The Immunohistochemical Localization of Fas and Fas Ligand in Jaw Bone and Tooth Germ of Human Fetuses

Abstract The cellular localization and roles of bone morphogenetic protein (BMP)-2 and apoptosis-associating factors in human orofacial development remain unclear. In this study, BMP-2, osteocalcin, and TGF-β, which are bone-differentiating markers, apoptosis-associating factors (i.e., Bcl-2, Bax, Fas, and Fas ligand), apoptotic cells detected by the in situ 3′-end labeling method (TUNEL), and proliferating cell nuclear antigen (PCNA) were immunohistochemically examined in the heads (in particular, the jaw bone and tooth germs) of human fetuses of 11-week pregnancy. BMP-2 was positive in osteoblasts and newly formed osteoid of the incisive and palatal bone of the maxilla and the mandible, which indicated that BMP-2 was exclusively involved in intramembranous ossification in the human fetal head. Fas was positive in the cytoplasm of osteocytes and a few osteoblasts. In contrast, Fas ligand was positive in the cytoplasm of osteoblasts and abundant in the stroma of the osteoblastic layer, periosteum, and perichondrium. The Fas ligand in the stroma was recognized as the soluble form, which was possibly produced by osteoblasts. TUNEL-positive apoptotic cells were found in a few osteocytes and a few osteoblastic cells in new bone, and in monocytes of degenerate Meckels cartilage. The induction of apoptosis observed in monocytes seems to be caused via a Fas-Fas ligand cell death system, because some of these monocytes were Fas-positive, and most of them were Fas ligand-positive. Interestingly, the abundant soluble Fas ligand observed in the periosteum probably protects the bone-formative zone from the invasion of the activated lymphocytes by binding to Fas expressing in these lymphocytes and killing these cells. Fas and Fas ligand were focally positive in the dental lamina and inner enamel epithelium and cusps of the enamel organ, nevertheless, the presence of TUNEL-positive cells was very rare. Bcl-2 was clearly and Bax was weakly positive in the cells throughout the dental lamina and enamel organ. These findings indicated that Fas-mediated apoptosis was inhibited by the Bcl-2 family in the development of teeth.

Harderianization is another sexual dimorphism of rat exorbital lacrimal gland.

The exorbital lacrimal glands (ELG) of rats were examined for both sexes to determine what degree of harderianization occurred as a function of age and after castration, and to investigate its time course and origin in ELG. Light microscopically, very small Harderian foci were seen in the ELG of both sexes at 3 weeks of age. As the male rats became older, the relative volume of the Harderian gland (HG) cells in the ELG increased. At age 6 months, the value was 1.25 +/- 0.31% in males and 0.13 +/- 0.05% in females (p less than 0.05). After castration, a significant decrease (0.21 +/- 0.01%, p less than 0.05) was observed in that of male ELG. In contrast, in female ELG, HG cells were inconspicuous and the relative volume of those did not vary during this experimental period or after castration. It appeared that the HG cells had developed from undifferentiated basal cells of the acini and the intercalated ducts in the ELG at age 2-6 months. Then, at age 22 months, they also probably developed from those of the excretory ducts of the ELG.

A sexual dimorphism of mucous cells in the submandibular salivary gland of rat

Submandibular glands of both sexes from one week to six months-old were serially sectioned, and mucous cells showing a strong positive stain to alcian blue pH 2.5, first detected at two weeks, showed a high degree of inter-sectional and inter-individual variability. In males, the percentage of animals having mucous cells increased with sexual maturation and attained 100 per cent at age six months. In females, the maximum (60 per cent) was reached by four weeks and then decreased with sexual maturation. Only 28.5 per cent of the glands of adult female rats contained mucous cells. In the six-month-old females, the percentage of female rats having mucous cells increased three-fold after injection of testosterone (total dose 17-21 mg; p less than 0.05, Fisher test). Mucous cells in testosterone-injected females displayed morphological features which suggest a transition from original seromucous cells to mucous cells.

Gastric carcinoma with psammomatous calcification after Billroth II reconstruction : Case report and literature review

A case of gastric carcinoma with psammomatous calcification arising in the remnant stomach after Billroth II reconstruction is reported. Borrmann type 1 gastric carcinoma was detected in the remnant stomach of an 82‐year‐old woman, who had a past history of distal partial gastrectomy for a perforated gastric ulcer, with Billroth II reconstruction at 40 years of age. Histologically, the tumor was a tubular adenocarcinoma that invaded the muscularis propria. Numerous psammoma bodies were found in the lumens of the tumor glands. Dystrophic calcification of gastric cancer is rare and psammomatous calcification of gastric cancer has only been reported in five cases previously. To our knowledge, this is the first case of gastric carcinoma with psammomatous calcification arising in the remnant stomach. We also review previously published reports regarding gastric carcinoma with psammomatous calcification.

Esophageal undifferentiated carcinoma displaying marked chondroid differentiation at metastatic foci

A report of an unusual esophageal tumor in an 81‐year‐old man is presented. The primary tumor was diagnosed as undifferentiated carcinoma at biopsy and had disappeared after irradiation treatment. However, multiple metastases were noted in the brain, lungs, kidneys, adrenals and spleen at autopsy. Histologically, metastases showed marked cartilaginous metaplasia as demonstrated by light microscopy, histochemical and immunohistochemical studies, although the initial biopsy sample did not possess chondroid matrix. Furthermore, an apparent transition could be traced from carcinomatous to chondroid cells, suggesting that the chondroid cells were derived from carcinoma cells. The carcinomatous area partially showed both squamous and glandular differentiation, although they were poorly differentiated. A retrospective immunohistochemical study that used a panel of antibodies suggested a phenotypic relevance between primary and metastatic tumors.

Glucocorticoid-induced growth inhibition with enhanced expression of ductal epithelium of human salivary gland adenocarcinoma cells transplanted into athymic nude mice.

The effect of glucocorticoid on growth and the histologic features of adenocarcinoma induced in nude mouse by the inoculation of neoplastic epithelial cells of salivary duct origin (HSG) were studied. Subcutaneous injection of dexamethasone (low‐dose group: 0.05 μg/g, high‐dose group: 0.25 μg/g) four times a week for 8 weeks significantly (P < 0.05) inhibited tumor growth, and in one mouse the tumor had almost completely disappeared by the middle of the sixth week of treatment. In addition, dexamethasone induced an apparent luminal structure in the tissue section of the tumor and enhanced the immunoperoxidase reaction to epithelial membrane antigen. The results indicate that dexamethasone inhibited the proliferation of HSG cells in the nude mouse transplantation system probably by inducing the cellular differentiation of the HSG cells toward the more differentiated ductal epithelia via glucocorticoid receptors.

ULTRASTRUCTURAL STUDY OF BENIGN OSTEOBLASTOMA OF THE MAXILLA

The ultrastructure of benign osteoblastoma of the maxilla in a 14‐year‐old boy was studied. Morphologically, the tumor tissue was composed of abundant osteoid of a trabecular form, and cellular and vascular connective tissue. The cellular components were osteoblasts, osteoclasts, osteocytes entrapped within the osteoid, and small perivascular cells of two types. The small cells of one type were ovoid and had a relatively large nucleus and a dark cytoplasm. The cells of the other type were elliptical and had a clear cytoplasm. The former seemed to be a preosteoblast and the latter could not be identified. The osteoblast was the predominant cell in this lesion and was characterized by the presence of abundant rough‐surfaced endoplasmic reticulum and several Golgi complexes. These characteristics indicated the pronounced activity of collagenous fiber synthesis and the matrix of the osteoid tissue.

Esophageal squamous cell carcinoma characterized by extensive chondroid differentiation

A rare case of carcinoma characterized by extensive chondroid elements at a site of primary esophageal and metastatic lesion is reported. The patient was a 67‐year‐old man complaining of dysphagia due to an ulcerative lesion at the lower middle esophagus. He underwent irradiation treatment prior to surgery. Histologically, the tumor consisted of both carcinomatous and chondroid elements and had invaded deeply into the esophageal wall. The carcinomatous cells had gradually become chondroid cells embedded within an extensive extracellular matrix. In addition, the metastatic lesion showed findings similar to those of the primary lesion. Immunohistochemistry revealed that both carcinomatous and chondroid elements were immunostained with cytokeratin and epithelial membrane antigen, suggesting an epithelial nature to the chondroid cells. Conversely, only chondroid cells were positively stained for S‐100 protein. Furthermore, bone morphogenetic proteins (BMP) were positive for chondroid cells and their surrounding carcinomatous cells. Given the apparent transition between carcinomatous and chondroid cells based on microscopy and immunohistochemical findings in the present case, we concluded that the chondroid cells were derived from carcinomatous cells. In addition, our findings suggest that BMP produced by carcinomatous cells lead to chondroid differentiation of the carcinoma cells.