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Establishment and characterization of a novel gastric cancer cell line from dermatomyositis patient
406
REGULATORY PIECHANISIJSOF EXPRESSION OF A HUMAN GENE ENCODING FOR THE MELANOMA-ASSOCIATED ME491 ANTIGEN. HAK HOTTA. Department of Microbiology, of Medicine, Kobe, Hyogo.
Kobe University
School
A
human gene encoding for a melanoma-associated antigen was molecularly cloned from a human library, and regulatory mechanisms of the genomic gene expression was analyzed. The promoter, identified by nucleotide sequencing and primer extension was found to utilize the GC box but not analyses, the TATA box. CAT assay revealed that the promoter cells exerted its activity in Ha-ras-transformed more efficiently than in non-transformed cells. It was also found that a cryptic promoter sequence in ME491 antigen an intron, which could initiate expression, was partially repressed by an upstream repressor sequence. These positive and negative for the regulatory sequences may be responsible stage-dependent expression of ME491 antigen. ME491
CYTOFLUOROMETRICAL ANALYSISOF DNA HISTOGRAMOF PAGET CELLS IN EXTRAMAMMA RY PAGET'S DISEASE. OSAMU MORI, HIROSHI HACHISUKA, AND YOICHIRO SASAI.
Departmentof Dermatology,Kurume Univ. School of Medicine, Kurume. Separationof Paget cells was performedfrom five cases of extramammary Paget's disease. Stripped skin was treated with EDTA and trypsin, and epidermal cell suspension was obtained. No dermal invasion and metastasis of regional lymph nodes was seen in four cases. In a case, in which a thumb sized red nodule was seen, dermal invasion and metastasis of regional lymph nodes were present. DNA histogram of Paget's cells revealed biphasic pattern in the four cases, whereas the latter case where the nodule was present showed aneuploid pattern. The aneuploid patternwas seen in Paget cells from erythematouslesion as well as in Paget cells from the nodule. The aneuploid found in DNA histogramindicatesthe biological behavior of PaSet cells.
ESTABLISHMENT AND CHARACTERIZATION OF A NOVEL GASTRIC CANCER CELL LINE FROM DERMATOMYOSITIS PATIENT. ATSUKO SEKINE. HIKARU ETO. YUKINORI OHTA. AND SHIGEO NISHIYAMA. Department of Dermatology, Kitasato University School of Medicine, Sagamihara. We have established a novel gastric cancer cell line, MKYT. from surgically removed gastric cancer tissue of 58 y/o dermatomyositis patient. The cells are growing in RPM1 1640 medium supplemented with 10% FBS in stable condition for more than 1 year. The doubling time of MKYT cells is about 34 hours. Chromosome analysis of these cells revealed modal number of 43 with some marker chromosomes. The tumor cells grew rapidly in nude mice, however, none of the mice developed myositis. Surprisingly, when culture supernatant from MKYT cells was added to rat muscle cell (L6) culture, the growth of L6 was markedly increased, suggesting that this cell line produces muscle cell growth factor in vitro. Control gastric cancer cell line did not show such activity. MKYT cell line would be useful tool to study pathogenesis of dermatomyositis. WHY CAN EPIDERMAL CYSTS BE EASILY PULLED OUT? YOSHIHIKD MITSUHASHI, NAOMI AKITA AND YUKO TAKAGI Department of Dermatology,Hirosaki University School of Medicine, Hirosaki Non-infected epidermal cysts can easily be pulled out. The above clinical observation can be supported by the histopathological finding that the wall of epidermal cysts are frequently separated from the dermal matrix. Why are they easily separated from the dermis? Two explanations are possible. First, their adherence system of the dermal-epidermaljunction area is impaired. Second, sorrounding dermal matrix is altered. We examined epidermal cysts by inniunopathological techniques focusing on the dermal-epidermaljunction area of the cvsts, and compared it with that of the eaidermis. We studied BP antigen; laminin, type IV and type Vii collagen. We have also done an electron microscopical observation. The results showed that all the observed components of the epidermal cysts are the same as those of the epidermis. We coneluded that the easiness of separation may be contributed to alternations surrounding dermal matrix.
ULTRASTRUCTUREOF THE LESIONS OF EXPERIMENTAL SKIN CANCER AFTER TOPICAL APPLICATION OF GLUTARALDEHYDESOLUTION
IMMUNOHISCAL CHARACTERIZATION OF EPIDERMAL PROTEINS IN CALCIPYINGEPITHBLIOMA
KEIICHI UEDA, KAZUMORI ISHIGURO AND KEN-ICHI KAWAHARA Department of Dermatology, Fukui Medical School, Fukui, Japan
SHINICHI WATANABE, KBIKO WAGATSUMA, EIKO ICHIKAWA, MASAKO MIZGGUCHI, AND HISASIB TAKAHASHI. Department of Dermatology, Teikyo University School of Medicine, Tokyo, Japan. The present study was conducted to examine the immunohistochemical distributionof various epidermat proteins in calcifying epithelioma (CA) in order to confbm the histogenesis of this tumor. Specimens were obtained from 15 cases of CA. Various kinds of anti-cytokeratin antibodies, anti-humanfilaggrin antibody, and rabbit amiserum against involucrin were used as primary antibodies. Stainings were performed using the avidin-biotin immunoperoxidasemedmd. In CA, only transitionalcelts were strongly stained with involuctin, while basophilic cells and shadow cells showed no reactivity with involucrin or the other cpidermal protein antibodies. Epidermoid cells were stained with some anti-keratinantibodies. In normal hair, the inner root sheath reveated positive stainings with only No.13 cytokeratin and involucrin antibodies. The hair medulla and the lower part of the hair cortex expressed only involucrin. These findings suggest that the tmnor arises from hair germ cells which are differentiating toward hair cortex. The transitional cells are assumed to cormspond to the lower hair cortex and the shadow cells to keratinous xone of the hair.
Ultrastructural investigationwas performed on the lesions of experimental skin cancer in C3H mice. 0.5 ml of 2% glutaraldehyde solution was injected to the lesions. Twenty four hours after injection, the lesions were biopsied, and ultrastructural investigationwas performed. Nuclei of the tumor cells were pyknotic and nucleoplasm was uneven in figure. In the cytoplasm, vacuoles were increased in number, and mitochondria were swollen. Some of them showed spongelike configuration. Capillaries were filled with erythrocytes and vacuoles were increased in the cytoplasm of endothelial cells. These findings showed that glutaraldehyde solution was effective against the lesions of experimental skin cancer.
REGULATORY PIECHANISIJSOF EXPRESSION OF A HUMAN GENE ENCODING FOR THE MELANOMA-ASSOCIATED ME491 ANTIGEN. HAK HOTTA. Department of Microbiology, of Medicine, Kobe, Hyogo.
Kobe University
School
A
human gene encoding for a melanoma-associated antigen was molecularly cloned from a human library, and regulatory mechanisms of the genomic gene expression was analyzed. The promoter, identified by nucleotide sequencing and primer extension was found to utilize the GC box but not analyses, the TATA box. CAT assay revealed that the promoter cells exerted its activity in Ha-ras-transformed more efficiently than in non-transformed cells. It was also found that a cryptic promoter sequence in ME491 antigen an intron, which could initiate expression, was partially repressed by an upstream repressor sequence. These positive and negative for the regulatory sequences may be responsible stage-dependent expression of ME491 antigen. ME491
CYTOFLUOROMETRICAL ANALYSISOF DNA HISTOGRAMOF PAGET CELLS IN EXTRAMAMMA RY PAGET'S DISEASE. OSAMU MORI, HIROSHI HACHISUKA, AND YOICHIRO SASAI.
Departmentof Dermatology,Kurume Univ. School of Medicine, Kurume. Separationof Paget cells was performedfrom five cases of extramammary Paget's disease. Stripped skin was treated with EDTA and trypsin, and epidermal cell suspension was obtained. No dermal invasion and metastasis of regional lymph nodes was seen in four cases. In a case, in which a thumb sized red nodule was seen, dermal invasion and metastasis of regional lymph nodes were present. DNA histogram of Paget's cells revealed biphasic pattern in the four cases, whereas the latter case where the nodule was present showed aneuploid pattern. The aneuploid patternwas seen in Paget cells from erythematouslesion as well as in Paget cells from the nodule. The aneuploid found in DNA histogramindicatesthe biological behavior of PaSet cells.
ESTABLISHMENT AND CHARACTERIZATION OF A NOVEL GASTRIC CANCER CELL LINE FROM DERMATOMYOSITIS PATIENT. ATSUKO SEKINE. HIKARU ETO. YUKINORI OHTA. AND SHIGEO NISHIYAMA. Department of Dermatology, Kitasato University School of Medicine, Sagamihara. We have established a novel gastric cancer cell line, MKYT. from surgically removed gastric cancer tissue of 58 y/o dermatomyositis patient. The cells are growing in RPM1 1640 medium supplemented with 10% FBS in stable condition for more than 1 year. The doubling time of MKYT cells is about 34 hours. Chromosome analysis of these cells revealed modal number of 43 with some marker chromosomes. The tumor cells grew rapidly in nude mice, however, none of the mice developed myositis. Surprisingly, when culture supernatant from MKYT cells was added to rat muscle cell (L6) culture, the growth of L6 was markedly increased, suggesting that this cell line produces muscle cell growth factor in vitro. Control gastric cancer cell line did not show such activity. MKYT cell line would be useful tool to study pathogenesis of dermatomyositis. WHY CAN EPIDERMAL CYSTS BE EASILY PULLED OUT? YOSHIHIKD MITSUHASHI, NAOMI AKITA AND YUKO TAKAGI Department of Dermatology,Hirosaki University School of Medicine, Hirosaki Non-infected epidermal cysts can easily be pulled out. The above clinical observation can be supported by the histopathological finding that the wall of epidermal cysts are frequently separated from the dermal matrix. Why are they easily separated from the dermis? Two explanations are possible. First, their adherence system of the dermal-epidermaljunction area is impaired. Second, sorrounding dermal matrix is altered. We examined epidermal cysts by inniunopathological techniques focusing on the dermal-epidermaljunction area of the cvsts, and compared it with that of the eaidermis. We studied BP antigen; laminin, type IV and type Vii collagen. We have also done an electron microscopical observation. The results showed that all the observed components of the epidermal cysts are the same as those of the epidermis. We coneluded that the easiness of separation may be contributed to alternations surrounding dermal matrix.
ULTRASTRUCTUREOF THE LESIONS OF EXPERIMENTAL SKIN CANCER AFTER TOPICAL APPLICATION OF GLUTARALDEHYDESOLUTION
IMMUNOHISCAL CHARACTERIZATION OF EPIDERMAL PROTEINS IN CALCIPYINGEPITHBLIOMA
KEIICHI UEDA, KAZUMORI ISHIGURO AND KEN-ICHI KAWAHARA Department of Dermatology, Fukui Medical School, Fukui, Japan
SHINICHI WATANABE, KBIKO WAGATSUMA, EIKO ICHIKAWA, MASAKO MIZGGUCHI, AND HISASIB TAKAHASHI. Department of Dermatology, Teikyo University School of Medicine, Tokyo, Japan. The present study was conducted to examine the immunohistochemical distributionof various epidermat proteins in calcifying epithelioma (CA) in order to confbm the histogenesis of this tumor. Specimens were obtained from 15 cases of CA. Various kinds of anti-cytokeratin antibodies, anti-humanfilaggrin antibody, and rabbit amiserum against involucrin were used as primary antibodies. Stainings were performed using the avidin-biotin immunoperoxidasemedmd. In CA, only transitionalcelts were strongly stained with involuctin, while basophilic cells and shadow cells showed no reactivity with involucrin or the other cpidermal protein antibodies. Epidermoid cells were stained with some anti-keratinantibodies. In normal hair, the inner root sheath reveated positive stainings with only No.13 cytokeratin and involucrin antibodies. The hair medulla and the lower part of the hair cortex expressed only involucrin. These findings suggest that the tmnor arises from hair germ cells which are differentiating toward hair cortex. The transitional cells are assumed to cormspond to the lower hair cortex and the shadow cells to keratinous xone of the hair.
Ultrastructural investigationwas performed on the lesions of experimental skin cancer in C3H mice. 0.5 ml of 2% glutaraldehyde solution was injected to the lesions. Twenty four hours after injection, the lesions were biopsied, and ultrastructural investigationwas performed. Nuclei of the tumor cells were pyknotic and nucleoplasm was uneven in figure. In the cytoplasm, vacuoles were increased in number, and mitochondria were swollen. Some of them showed spongelike configuration. Capillaries were filled with erythrocytes and vacuoles were increased in the cytoplasm of endothelial cells. These findings showed that glutaraldehyde solution was effective against the lesions of experimental skin cancer.