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130 p53 Gene mutations in porokeratosis and squamous cell carcinoma arising in porokeratosis
84
JSID Abstracts/ Journal of Dertnatological Science 10 (1995) 61-102
127 LIGHT
130 AND ELECTRON
PRIMARY
CULTURE
OF ECCRINE
Department of Dermatology, Tumor carcinoma kinetics.
cells
MICROSCOPIC
obtained
STUDIES
ON THE
DUCT CARCINOMA.
11
Fukui Medical School, Fukui, Japan. from
a 7%year-old
man with
eccrine
were cultured and were investigated their morphology The tumor tissues were transplanted
mice and then maintained for two weeks. mice were cut into I mm3 blocks
into subcutanea
duct
and cell of nude
The tumor tissues from nude
and the specimens were cultured
in
dishes (35 mm in diameter) using with minimum essential medium added 20 96 fetal bovine serum. cultured
Two and four weeks later, we examined the
tumor cells light and electron microscopically.
characteristics
of these tumor cells before cultivation
Morphological
were compared with
after cultivation.
128 FURTHER STIJD1E.SOF BIOLOGICAL AND Bl CCHFMICAL cHARACTRIsTIcs OF A G-CSF pROD”ClNG ECCltlNE ADENGCAKClNOMA CELL UNEL&&, T. Ksneksm. M. S~ovsms. sad T I@~&Depmimea~t of Damatdogy. Kagotima University Faculty of Medidne.Ka&ima. Japan. E&line carcinomacell cHained rrom a patientwho showedmarLedperipheral“elmopbilia was *Itabliskd in culntrc. Cell* have beenmaintainedin Eagle’sMl34 supplemamd withlO% FBS for 20 month.9with more &an 62 psgpag*l. Cells in culture produceda large ammmtof G-CSF, i& < 0. lag /ml ( bef-xe ). 1.4ng I ml ( growing ). and 26.5ng /ml (cmdluent phase).Injectiws of culturedmedia (25ml) into rabbit veins induced marked nsuvophilia (4 folds mwe thannormal). Cells were haasplantedinto atbymic nudemice and SCID mice. Marked mutrophilia WQSala0 observed in tumorbearing SCID mice in pqcxtion to the size.& 2.1001mmyh2rm tmnsplanralial). 4o.OOO/mn?(lcm-tumor), 12O.C0O/mm’(24 and 2OO.OWmm9(3cm) Aftu tumor resectionscutmpbil number8inathymicnudemiceR4Ured tonamal.~.l5O.@Xl/~ (3a,wmor). 67,000/mm’(7days alter IEUC&OO).sud 5.8OWmn?(28days). G-CSF WBS The exprmiw of immunobisWcally detectedio the cytcplasm of tumor cell. mesmger RNA OTG-CSF wa, detectedin wltured calls by KT-Xl3 m&cd. and it WBS sot &tecIed in other cell lises.~ .KHm-l~mdaaoma), K-TL..l(aichilemmass), and K-MFH-l(malignaat tibmw histionna).Na~iphilia with the p~timt was mntimwd to be cawed by G-CSF prwkced by emine cradmma cells.
p53 GENE MUTATIONS IN POROKERATOSIS AND SQUAMOUS CELL CAAClNOMAARlSlNG IN POROKEFLATOSIS. Y. NINOMIYA’. S SASAKI. Y. KUBO. Y. URANO. S. ARASE. De+mrtrnsnt of Dsrntetology, ThsUliiersity of Tokushima, &hod of M&tine, Tokushima, Japan. Skin csnsometimes develop in pomkeraotii iedons. Mdecular mechanisms of the skin carcinogenesis remain to bs duddated. The p53 tumor suppressor gsne is most commonly mutated in vatious human tumors induding skin cancers and precancerous lesions of the skin. We previously reported lhst immunohiitodmmical p53 oversxpresskm was frequently detected in pxokeratotic &ions and SCCs arising in pcmketatosis. In thii study, to sse whether p53 gene mutations oxrelate to the etiology of porokeratosis and the aubae quent csrcinogsnesis. poroksratotic lesions and SC& arising in purokeretosis were examined on the mutations by pdymsrass chain reactiin-single strand conformation polymorphism (PC&SSCP) analysis. A total of 24 DNA samples (18 01poroksratosis and 6 01SCC) were obtained from 9 patients induding 4 of the plaque type. 3 of the disseminated superficial actinic pofoksratosis, one 01 the giant type and one of the linear type. Of the 6 SCC DNA& live were prepared from a patient with linear pofokerstosis with muiliple SCCs. and one was prepared from a patient wilh giant porokeratosis. PCR-SSCP analyses in sxon 5 to 10 01 the p53 gene showed abnormal bands in one 01 the 18 porokerstotic DNAs and two of the 6 SCC DNAs. All three DNAs showing mobility shit were from the patient with linear poroksratosis. We are now trying to determine nucleotids sequences of the three DNAs to confirm the presence of ~53 gene mutations. Our findings suggest that p53 gene mutations are not essential in poroksratosis but have some relevance to the csrcinogenssis of porokeratosis.
131
in ttm win includinIJcTcL, lyqhpmlifrrativa diCBCL, lti-l lymplxu~, lymphmatoid papllais, lywhdmumia bmnign.6 cuti* aa lymghmqt ic infiltration Of ttw mkin. A few ZAQoptotic calls famdincu-*l~. whih n-rcu. apogtotic OnllO in tha l4pF.r dermim in the lemianal skin of letoid pa~~lcaia.
129
132
HISTOCHEMICAL DISTINCI’ION BETWEEN TRICHOLEMMAL CARCINOMA AND SQUAMO*US CELL CARCINOI?A OFTHE SKIN USING LECTIN UEA-I. uo . T. Murw Department of Dermatology, Nam Medical University, Nara Japan. Tricholemmal carcinoma (TLC) is a cutaneous appendage tumor. In some cases, histological distinction between TLC and squamous cell carcinoma (SCC) of the skin is dif2icuIt by H&E staining. We tried to establish the simple and reliable method to distinguish TLC from SCC. In the present study we examined the specimens of TLC (4 cases) and SCC of the skin (10cases)using the lectin Ulex europaeus agglutinin-I (UEA-I). In normal skin samples UEA-I wBs positive in the outer root sheath cells of the hair follicle, and upper spinous and granular layers of the epidermis. In all four cases of TLC, tumor cells were positively stained with UEA-I. On the other hand, tumor c&s of SCC showed negative stained with UEA-I. This result indicates that histochemical staining by using UEA-I is a simple and useful method for distinction between TLC and SCC of the skin.
EPSTEIN-BARR (EB) VIRUS-RELATED LYMPHOMAS: DETECTION OF EB VIRUS GENES USING THREE DIFFERENT METHODS. M. H. Dspaltment of Dermstdogy, Fukushima Medical Collage, Fukushima, Japan. We investigated the presence of EB virus genes and their pfoducts in various types of cutaneous lymphcmss, using PCR, in situ hybridization (ISH) and immunostsin for latent membrane protein (LMPl). Among 40 patients with cutaneous ~mpttomas or related dknrden, EB virus gene was detected in 2 cases. Their clinical diagnoses were of T-cell lymphoma with clinical features of cytophsgic histiocytic psnniculitis and B-cell lymphoma showing hsmophsgaytosk,
reapectivdy.
The ISH stt@ on the both
cases demonstrated E&encoded RNAs in most infiltrating cells, suggesting a latent EB virus infection. The immumxtain with anti LMP-1 ant&acty, however, wss ahlost negative. lhsse dsts suggsst that EB virus is involved in the development of certsin types of cutarwous
Iympbomas, and that both PCR and ISH metboda are
useful for the screening test for EB virus.
JSID Abstracts/ Journal of Dertnatological Science 10 (1995) 61-102
127 LIGHT
130 AND ELECTRON
PRIMARY
CULTURE
OF ECCRINE
Department of Dermatology, Tumor carcinoma kinetics.
cells
MICROSCOPIC
obtained
STUDIES
ON THE
DUCT CARCINOMA.
11
Fukui Medical School, Fukui, Japan. from
a 7%year-old
man with
eccrine
were cultured and were investigated their morphology The tumor tissues were transplanted
mice and then maintained for two weeks. mice were cut into I mm3 blocks
into subcutanea
duct
and cell of nude
The tumor tissues from nude
and the specimens were cultured
in
dishes (35 mm in diameter) using with minimum essential medium added 20 96 fetal bovine serum. cultured
Two and four weeks later, we examined the
tumor cells light and electron microscopically.
characteristics
of these tumor cells before cultivation
Morphological
were compared with
after cultivation.
128 FURTHER STIJD1E.SOF BIOLOGICAL AND Bl CCHFMICAL cHARACTRIsTIcs OF A G-CSF pROD”ClNG ECCltlNE ADENGCAKClNOMA CELL UNEL&&, T. Ksneksm. M. S~ovsms. sad T I@~&Depmimea~t of Damatdogy. Kagotima University Faculty of Medidne.Ka&ima. Japan. E&line carcinomacell cHained rrom a patientwho showedmarLedperipheral“elmopbilia was *Itabliskd in culntrc. Cell* have beenmaintainedin Eagle’sMl34 supplemamd withlO% FBS for 20 month.9with more &an 62 psgpag*l. Cells in culture produceda large ammmtof G-CSF, i& < 0. lag /ml ( bef-xe ). 1.4ng I ml ( growing ). and 26.5ng /ml (cmdluent phase).Injectiws of culturedmedia (25ml) into rabbit veins induced marked nsuvophilia (4 folds mwe thannormal). Cells were haasplantedinto atbymic nudemice and SCID mice. Marked mutrophilia WQSala0 observed in tumorbearing SCID mice in pqcxtion to the size.& 2.1001mmyh2rm tmnsplanralial). 4o.OOO/mn?(lcm-tumor), 12O.C0O/mm’(24 and 2OO.OWmm9(3cm) Aftu tumor resectionscutmpbil number8inathymicnudemiceR4Ured tonamal.~.l5O.@Xl/~ (3a,wmor). 67,000/mm’(7days alter IEUC&OO).sud 5.8OWmn?(28days). G-CSF WBS The exprmiw of immunobisWcally detectedio the cytcplasm of tumor cell. mesmger RNA OTG-CSF wa, detectedin wltured calls by KT-Xl3 m&cd. and it WBS sot &tecIed in other cell lises.~ .KHm-l~mdaaoma), K-TL..l(aichilemmass), and K-MFH-l(malignaat tibmw histionna).Na~iphilia with the p~timt was mntimwd to be cawed by G-CSF prwkced by emine cradmma cells.
p53 GENE MUTATIONS IN POROKERATOSIS AND SQUAMOUS CELL CAAClNOMAARlSlNG IN POROKEFLATOSIS. Y. NINOMIYA’. S SASAKI. Y. KUBO. Y. URANO. S. ARASE. De+mrtrnsnt of Dsrntetology, ThsUliiersity of Tokushima, &hod of M&tine, Tokushima, Japan. Skin csnsometimes develop in pomkeraotii iedons. Mdecular mechanisms of the skin carcinogenesis remain to bs duddated. The p53 tumor suppressor gsne is most commonly mutated in vatious human tumors induding skin cancers and precancerous lesions of the skin. We previously reported lhst immunohiitodmmical p53 oversxpresskm was frequently detected in pxokeratotic &ions and SCCs arising in pcmketatosis. In thii study, to sse whether p53 gene mutations oxrelate to the etiology of porokeratosis and the aubae quent csrcinogsnesis. poroksratotic lesions and SC& arising in purokeretosis were examined on the mutations by pdymsrass chain reactiin-single strand conformation polymorphism (PC&SSCP) analysis. A total of 24 DNA samples (18 01poroksratosis and 6 01SCC) were obtained from 9 patients induding 4 of the plaque type. 3 of the disseminated superficial actinic pofoksratosis, one 01 the giant type and one of the linear type. Of the 6 SCC DNA& live were prepared from a patient with linear pofokerstosis with muiliple SCCs. and one was prepared from a patient wilh giant porokeratosis. PCR-SSCP analyses in sxon 5 to 10 01 the p53 gene showed abnormal bands in one 01 the 18 porokerstotic DNAs and two of the 6 SCC DNAs. All three DNAs showing mobility shit were from the patient with linear poroksratosis. We are now trying to determine nucleotids sequences of the three DNAs to confirm the presence of ~53 gene mutations. Our findings suggest that p53 gene mutations are not essential in poroksratosis but have some relevance to the csrcinogenssis of porokeratosis.
131
in ttm win includinIJcTcL, lyqhpmlifrrativa diCBCL, lti-l lymplxu~, lymphmatoid papllais, lywhdmumia bmnign.6 cuti* aa lymghmqt ic infiltration Of ttw mkin. A few ZAQoptotic calls famdincu-*l~. whih n-rcu. apogtotic OnllO in tha l4pF.r dermim in the lemianal skin of letoid pa~~lcaia.
129
132
HISTOCHEMICAL DISTINCI’ION BETWEEN TRICHOLEMMAL CARCINOMA AND SQUAMO*US CELL CARCINOI?A OFTHE SKIN USING LECTIN UEA-I. uo . T. Murw Department of Dermatology, Nam Medical University, Nara Japan. Tricholemmal carcinoma (TLC) is a cutaneous appendage tumor. In some cases, histological distinction between TLC and squamous cell carcinoma (SCC) of the skin is dif2icuIt by H&E staining. We tried to establish the simple and reliable method to distinguish TLC from SCC. In the present study we examined the specimens of TLC (4 cases) and SCC of the skin (10cases)using the lectin Ulex europaeus agglutinin-I (UEA-I). In normal skin samples UEA-I wBs positive in the outer root sheath cells of the hair follicle, and upper spinous and granular layers of the epidermis. In all four cases of TLC, tumor cells were positively stained with UEA-I. On the other hand, tumor c&s of SCC showed negative stained with UEA-I. This result indicates that histochemical staining by using UEA-I is a simple and useful method for distinction between TLC and SCC of the skin.
EPSTEIN-BARR (EB) VIRUS-RELATED LYMPHOMAS: DETECTION OF EB VIRUS GENES USING THREE DIFFERENT METHODS. M. H. Dspaltment of Dermstdogy, Fukushima Medical Collage, Fukushima, Japan. We investigated the presence of EB virus genes and their pfoducts in various types of cutaneous lymphcmss, using PCR, in situ hybridization (ISH) and immunostsin for latent membrane protein (LMPl). Among 40 patients with cutaneous ~mpttomas or related dknrden, EB virus gene was detected in 2 cases. Their clinical diagnoses were of T-cell lymphoma with clinical features of cytophsgic histiocytic psnniculitis and B-cell lymphoma showing hsmophsgaytosk,
reapectivdy.
The ISH stt@ on the both
cases demonstrated E&encoded RNAs in most infiltrating cells, suggesting a latent EB virus infection. The immumxtain with anti LMP-1 ant&acty, however, wss ahlost negative. lhsse dsts suggsst that EB virus is involved in the development of certsin types of cutarwous
Iympbomas, and that both PCR and ISH metboda are
useful for the screening test for EB virus.