２２回日本研究皮膚科学会総会・抄録

２２回日本研究皮膚科学会総会・抄録

-74- Carcinogenesis/Tomor/Oncogene(3) 9/4 Room-D
RELATIONSHIP BETWEEN E-CADHERIN AND LYMPHNODE METASTASIS IN HUMAN SQUAMOUS CELL CARCINOMA OF THE SKIN.
Shin Koseki, Takehiko Aoki, Yutaka Hozumi, Yoshihiko Mitsuhashi, Shigeo Kondo.
Department of Dermatology, Yamagata University School of Medicine, Yamagata, Japan.
It is well known that E-cadherin expression has the tendency to decrease in human internal organ carcinomas with metastasis, as compared with those without metastasis. We examine immunohistochemically the expression of E-cadherin with anti E-cadherin monoclonal antibody , and compared primary lesions of human squamous cell carcino-ma of the skin (SCC) with lymphatic metastasis, with those without lymphatic metastasis. Fifty-seven cases of SCC, which were formalin-fixed and paraffin-embedded, were employed. E-cadherin was negative or decreased in the majority cases of SCC with lymphatic metastasis. Our result indicates that the expression of E-cadherin correlates with lymphatic metastasis in SCC, just as observed in internal organ carcinomas.

-75- Carcinogenesis/Tomor/Oncogene(3) 9/4 Room-D
AN IMMUNOHISTOCHEMICAL STUDY OF AN UROKINASE RECEPTOR IN ECCRINE POROCARCINOMA AND ECCRINE POROMA.
Junichi Mizushima, Tatsutoshi Nogita, Makoto Kawashima.
Department of Dermatology, Tokyo Women's Medical College, Tokyo, Japan.
Urokinase-type plasminogen activator (uPA) is a serine protease which converts the proenzyme plasminogen to the active enzyme plasmin. Through the binding to a specific cell surface receptor (uPAR), urokinase shows accelerated enzymatic activity and has been found to be associated with the invasion and metastasis by degrading the component of the extracellular matrix in several malignant tumors. Eccrine porocarcinoma (EPCa) is a malignant tumor, which usually arises from a long-standing eccrine poroma (EP) and shows various clinical aspects. In order to clarify the association of uPAR with metastatic properties of EPCa, we investigated immunohistochemically the expression of uPAR in seven cases of EPCa and eight cases of EP. Only the tumor cells of the edges of tumor nests were positive for uPAR in 6 out of 8 cases of EP and one primary tumor of EPCa. One primary tumor of EPCa arises from EP, two primary tumors of EPCa without metastasis invading deeply in the dermis and three tumors with metastasis were all positive for uPAR. Metastatic tumor cells in lymph node were positive in 3 out of 3. It is, thus, suggested that uPAR might play a role in the metastasis in EPCa.

-76- Carcinogenesis/Tomor/Oncogene(3) 9/4 Room-D
iMUNOHISTOCHEMICAL DIFFERENCES OF the TUMOR CELLS IN RELATION TO MALIGNANT POTENTIAL AMANG MAMMARY-, EXTRAMAMMARY-PAGET`S DISEASE AND PAGET CARCINOMA.
Atsushi Yaguchi,Kohzou Yonemoto,Minako Tatsuta.
Department of Dermatology,Kitasato University School of Medicine,Sagamihara, Japan.
The purpos of study is to investigate immunohistochemically the differencies of tumor cell characteristics among mammary(mPD)-,extramammary-Paget`s disease(ePD) and Paget carci- noma(PC).20 specimens examined here were obtained from our patients 2 mPD, 9 ePD and 5 PC.Antibodies used this study were CA15-3,CD9,C-erb B2,E-cadherin. In brief,CA15-3 was positively stained in tumor cells of 3 disease . Tumor cells of ePD completely reacted CD9-positive, but the cells proliferated into dermis in PC tended to show negative. C-erb B2 showed clearly positive in all specimens of mPD and PC, as against less than half in ePD. On the contrary, E-cadherin was almost negative in the cells of mPD and PC exceot for those invading into lymphoid vessels, while mostly positive in common ePD. These immunohisto-chemical findings revealed the same tendency of reactivity in both mPD and PC, as compared with common ePD. It may indicate that these antibodies appear to aplicable to estimate the malignant potential in Paget`s disease.

-77- Carcinogenesis/Tomor/Oncogene(3) 9/4 Room-D
EXPRESSION OF CEA IN CULTURED PAGET CELLS PURIFICATED BY A COMBINATION METHOD USING ANTI-EMA ANTIBODY, IMMUNOBEADS, AND NYCODENZ DENSITY GRADIENT.
Osamu Mori, Minoru Miyasato, Tadashi Karashima, Takashi Hashimoto.
Department of Dermatology, Kurume University School of Medicine,Kurume, Japan.
We previously reported that CEA positive cells did not increase when cultured as epidermal cells, not Paget cells alone, from the involved skin with extramammary Paget's disease. In this study we examined whether CEA positive cells truly decrease or expression of CEA in Paget cells decrease. Skin sample was obtained from the scrotum of a 79-year-old man with extramammary Paget's disease, then epidermal cells were cultured as previously reported. The procurement of Paget cells was carried out from cultured epidermal cells by combining an anti-EMA monoclonal antibody, immunobeads, and a non-toxic and non-ionic density gradient medium, Nycodenz. . The resulting cells were washed with PBS, then, cultured with Keratinocyte-SFM. The cultured cells were subcultured, however, CEA positive cells were not increased. This study suggested that the decrease of CEA positive cells was due to the decline of expression of CEA in Paget cells.

-78- Carcinogenesis/Tomor/Oncogene(3) 9/4 Room-D
EFFECTS OF INTERFERONeb ON GROWTH OF MALIGNANT MELANOMA CELL LINES.
Hiromi Okazawa, Tetsuo Nagatani, Hiroshi Nakajima.
Department of Dermatology, Yokohama City University, School of Medicine, Yokohama, Japan.
IFNβ inhibits growth of the malignant melanoma cell lines. In this study, we tested whether IFNβ modulates the transcription activity of cyclin dependent kinase inhibitors in the malignant melanoma cell lines. After incubation of cell lines with or without IFNβ for 1 or 24 hrs, the expression levels of p16 and p21 mRNA were semi-quantified by RT-PCR. We observed transient increase of p21 expression. Secondly, we investigated the effect of combined treatment with IFNβ and steroid hormone. Cell proliferation inhibitory ratio was evaluated by CytoLite method. The anti-proliferative effect of IFNβ was enhanced by treatment with steroid hormone.

-79- Carcinogenesis/Tomor/Oncogene(3) 9/4 Room-D
DIFFERENTIAL APOPTOTIC PATTERN INDUCED BY PHOTODYNAMIC THERAPY AND CISPLATIN IN HUMAN SQUAMOUS CELL CARCINOMA CELL LINE.
Yoshinari Matsumoto1, Yoshinao Muro1, Yasuhiko Tamada2.
1Department of Dermatology, Nagoya University School of Medicine, Nagoya, 2Department of Dermatology, Aich Medical University, Aichi, Japan.
We compared the apoptotic cell death induced by photodynamic therapy (PDT) or cisplatin in a squamous cell carcinoma cell line, HSC-2. HSC-2 cells, kindly provided by Dr.T.Kuroki, Tokyo University, was cultured in MEM containing 10 % calf serum.The cells were treated with 3.0 μg/ml Photofrin II by 2.25 J/cm2 of UVA irradiation. They were also treated with 60 μg/ml of cisplatin. Total DNA was extracted and analyzed electrophoretically on a 2 % agarose gel. Nuclear matrix protein (NMP) in the culture supernatant was measured usining an enzyme immunoassay. NMP can be released from dying cells in a soluble form. PDT induced DNA fragmentation in oligonucleosome size within 2 h after PDT, whereas cisplatin 8 h after treatment. The release of NMP from HSC-2 cells was rapidly increased from 1 h after PDT, but no increase was detected within 12 h in cisplatin treatment. The nuclei were clearly seen in apoptotic cells induced by PDT, but were condensed in those induced by cisplatin. Differences on the HSC-2 cells were observed between the apoptosis induced by PDT and cisplatin. The mechanism for inducing apoptosis by PDT might be different from that by cisplatin.