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Dowling-Degos disease is genetically and clinico-pathologically distinct from reticulate acropigmentation of Kitamura, further confirmation
Abstracts / Journal of Dermatological Science 84 (2016) e1–e88
BRAF-activating somatic mutations often exist in malignant melanoma, however, the underlying molecular mechanism of somatic BRAF-mutation inductions remained to be clear. To explore the correlation between the AID expression and BRAF mutations, we performed the immunohistochemical study for 16 Japanese melanoma patients. Nine out of 10 malignant melanoma specimens with high AID expression in our study contained BRAFV600E mutation, while none of the 6 malignant melanomas with low AID expression had BRAFV600E mutation. Eight out of 10 malignant melanoma patients with high AID expression developed multi-organ metastases and/or multiple lymph node metastases afterwards during the follow-up, while no metastasis has been observed in melanoma patients with low AID expression. To further examine a role of AID in melanomas, we stably transfected normal human melanocytes with plasmids expressing AID under CMV promoter. Stable transfectant clones with high AID expression were selected with G418 and cultured with or without UVB irradiations for 150 days. Sequence analysis revealed the existence of BRAFV600E mutations in two independent clones with forced expression of AID and UV irradiations. Taken together, these data suggested that AID combined with other factors might play roles in inducing BRAFV600E mutation and can be a new target for prevention of malignant melanoma development. http://dx.doi.org/10.1016/j.jdermsci.2016.08.248 P13-04[C09-02] Suppressor of cytokine signaling-1 inhibits melanoma cell growth by the suppression of JAK/STAT and the activation of p53 signaling pathways Naoko Tagami 1,2,∗ , Satoshi Serada 2 , Minoru Fujimoto 2 , Atsushi Tanemura 1 , Tetsuji Naka 2 , Ichiro Katayama 1 1 Department of Dermatology, Osaka University Graduate School of Medicine, Japan 2 laboratory for Immuno Signal, National Institute of Biomedical Innovation, Osaka, Japan
Background: Melanoma is the most aggressive form of skin cancer, responsible for the majority of skin cancer related deaths. JAK-STAT signal is one of the main pathways involved in the tumor growth in melanoma. The suppressor of cytokine signaling (SOCS) family proteins are important negative regulator of proinflammatory cytokine signals by suppressing JAK-STAT signal pathway via inhibiting JAK activity. The aim of this study was to evaluate the role of SOCS-1 in the proliferation of melanoma and to clarify the mechanisms of its antitumor effect. Material and methods: Adenovirus vectors encoding SOCS1 gene was used to overexpress SOCS1 in G361, SK-MEL5, and SKMEL28 melanoma cell lines. The status of activated kinases and downstream proteins were determined by Western blot analysis. Anti-proliferative effects in melanoma cells were assessed by MTT assay. Apoptosis in melanoma cells was assessed by Caspase 3 activity and TUNEL staining. ICR nu/nu mice were used for subcutaneous xenograft experiment. Results: Overexpression of SOCS1 significantly reduced cell proliferation and induced apoptosis in G361 and SK-MEL5 not in SK-MEL28. Furthermore, in G361 and SK-MEL5 xenograft model, intratumoral injection of AdSOCS1 vector significantly reduced tumor volumes compared to AdLacZ-injected mice. Western blot and immunohistochemical analysis indicated that overexpression of SOCS1 induced apoptosis. The anti-proliferative effect of SOCS1
e81
correlated with decreased levels of the activation of STAT3 and increased levels of p53. Conclusions: We demonstrated that overexpression of SOCS-1 induced apoptosis in G361 and SK-MEL5 cells. We hope that these findings may lead to the successful clinical application of SOCS1 for melanoma treatment. http://dx.doi.org/10.1016/j.jdermsci.2016.08.249 P13-05[C09-03] Dowling-Degos disease is genetically and clinico-pathologically distinct from reticulate acropigmentation of Kitamura, further confirmation Michihiro Kono 1,∗ , Mutsumi Suganuma 1 , Hiromichi Takama 2 , Tamio Suzuki 3 , Kayoko Matsunaga 4 , Yasushi Tomita 1 , Masashi Akiyama 1 1 Department of Dermatology, Nagoya University Graduate School of Medicine, Nagoya, Japan 2 Takama Dermatology Clinic, Kasugai, Japan 3 Department of Dermatology, Yamagata University Faculty of Medicine, Yamagata, Japan 4 Department of Dermatology, Fujita Health University School of Medicine, Toyoake, Japan
Reticulate acropigmentation of Kitamura (RAK) [MIM# 615537] is a rare genetic disorder of cutaneous pigmentation with an autosomal dominant inheritance trait. The characteristic skin lesions are reticulate, slightly depressed brown macules mainly affecting the dorsa of the hands and the feet, which first appear before puberty and subsequently expand to the proximal limb and the trunk. On the other hand, Dowling-Degos disease (DDD) shows the similar skin manifestation of reticulate, slightly depressed brown macules on flexor regions of the extremities, mainly reported in Europe. There have been a controversy whether RAK and DDD were two distinct clinical entities or just on a spectrum of a single disease. We have reported that ADAM10 mutations cause RAK in 2013 (Kono et al, Hum Mol Genet 2013). In our previous report, we performed exome sequencing in a Japanese pedigree with RAK. A heterozygous mutation, [c.415C > T + c.424-425insCAGAG] (p.Pro139Ser + p.Arg142fsX43) in ADAM10 encoding a zinc metalloprotease, a disintegrin and metalloprotease domain-containing protein 10 (ADAM10) was identified in the RAK family. ADAM10 is known to be involved in the ectodomain shedding of various substrates in the skin. Sanger sequencing of ADAM10 in 5 more patients from 4 unrelated Japanese families identified 4 different mutations in ADAM10. Recently POFUT1 gene was reported as one of the causative genes of DDD. Thus, we performed mutation search for POFUT1 in one patient who showed atypical skin manifestation as RAK. In this paper, we report that a POFUT1 mutation, but no ADAM10 mutation was detected in the patient, which makes further confirmation that RAK and DDD are genetically and clinicopathologically independent disorders distinct from each other. http://dx.doi.org/10.1016/j.jdermsci.2016.08.250
BRAF-activating somatic mutations often exist in malignant melanoma, however, the underlying molecular mechanism of somatic BRAF-mutation inductions remained to be clear. To explore the correlation between the AID expression and BRAF mutations, we performed the immunohistochemical study for 16 Japanese melanoma patients. Nine out of 10 malignant melanoma specimens with high AID expression in our study contained BRAFV600E mutation, while none of the 6 malignant melanomas with low AID expression had BRAFV600E mutation. Eight out of 10 malignant melanoma patients with high AID expression developed multi-organ metastases and/or multiple lymph node metastases afterwards during the follow-up, while no metastasis has been observed in melanoma patients with low AID expression. To further examine a role of AID in melanomas, we stably transfected normal human melanocytes with plasmids expressing AID under CMV promoter. Stable transfectant clones with high AID expression were selected with G418 and cultured with or without UVB irradiations for 150 days. Sequence analysis revealed the existence of BRAFV600E mutations in two independent clones with forced expression of AID and UV irradiations. Taken together, these data suggested that AID combined with other factors might play roles in inducing BRAFV600E mutation and can be a new target for prevention of malignant melanoma development. http://dx.doi.org/10.1016/j.jdermsci.2016.08.248 P13-04[C09-02] Suppressor of cytokine signaling-1 inhibits melanoma cell growth by the suppression of JAK/STAT and the activation of p53 signaling pathways Naoko Tagami 1,2,∗ , Satoshi Serada 2 , Minoru Fujimoto 2 , Atsushi Tanemura 1 , Tetsuji Naka 2 , Ichiro Katayama 1 1 Department of Dermatology, Osaka University Graduate School of Medicine, Japan 2 laboratory for Immuno Signal, National Institute of Biomedical Innovation, Osaka, Japan
Background: Melanoma is the most aggressive form of skin cancer, responsible for the majority of skin cancer related deaths. JAK-STAT signal is one of the main pathways involved in the tumor growth in melanoma. The suppressor of cytokine signaling (SOCS) family proteins are important negative regulator of proinflammatory cytokine signals by suppressing JAK-STAT signal pathway via inhibiting JAK activity. The aim of this study was to evaluate the role of SOCS-1 in the proliferation of melanoma and to clarify the mechanisms of its antitumor effect. Material and methods: Adenovirus vectors encoding SOCS1 gene was used to overexpress SOCS1 in G361, SK-MEL5, and SKMEL28 melanoma cell lines. The status of activated kinases and downstream proteins were determined by Western blot analysis. Anti-proliferative effects in melanoma cells were assessed by MTT assay. Apoptosis in melanoma cells was assessed by Caspase 3 activity and TUNEL staining. ICR nu/nu mice were used for subcutaneous xenograft experiment. Results: Overexpression of SOCS1 significantly reduced cell proliferation and induced apoptosis in G361 and SK-MEL5 not in SK-MEL28. Furthermore, in G361 and SK-MEL5 xenograft model, intratumoral injection of AdSOCS1 vector significantly reduced tumor volumes compared to AdLacZ-injected mice. Western blot and immunohistochemical analysis indicated that overexpression of SOCS1 induced apoptosis. The anti-proliferative effect of SOCS1
e81
correlated with decreased levels of the activation of STAT3 and increased levels of p53. Conclusions: We demonstrated that overexpression of SOCS-1 induced apoptosis in G361 and SK-MEL5 cells. We hope that these findings may lead to the successful clinical application of SOCS1 for melanoma treatment. http://dx.doi.org/10.1016/j.jdermsci.2016.08.249 P13-05[C09-03] Dowling-Degos disease is genetically and clinico-pathologically distinct from reticulate acropigmentation of Kitamura, further confirmation Michihiro Kono 1,∗ , Mutsumi Suganuma 1 , Hiromichi Takama 2 , Tamio Suzuki 3 , Kayoko Matsunaga 4 , Yasushi Tomita 1 , Masashi Akiyama 1 1 Department of Dermatology, Nagoya University Graduate School of Medicine, Nagoya, Japan 2 Takama Dermatology Clinic, Kasugai, Japan 3 Department of Dermatology, Yamagata University Faculty of Medicine, Yamagata, Japan 4 Department of Dermatology, Fujita Health University School of Medicine, Toyoake, Japan
Reticulate acropigmentation of Kitamura (RAK) [MIM# 615537] is a rare genetic disorder of cutaneous pigmentation with an autosomal dominant inheritance trait. The characteristic skin lesions are reticulate, slightly depressed brown macules mainly affecting the dorsa of the hands and the feet, which first appear before puberty and subsequently expand to the proximal limb and the trunk. On the other hand, Dowling-Degos disease (DDD) shows the similar skin manifestation of reticulate, slightly depressed brown macules on flexor regions of the extremities, mainly reported in Europe. There have been a controversy whether RAK and DDD were two distinct clinical entities or just on a spectrum of a single disease. We have reported that ADAM10 mutations cause RAK in 2013 (Kono et al, Hum Mol Genet 2013). In our previous report, we performed exome sequencing in a Japanese pedigree with RAK. A heterozygous mutation, [c.415C > T + c.424-425insCAGAG] (p.Pro139Ser + p.Arg142fsX43) in ADAM10 encoding a zinc metalloprotease, a disintegrin and metalloprotease domain-containing protein 10 (ADAM10) was identified in the RAK family. ADAM10 is known to be involved in the ectodomain shedding of various substrates in the skin. Sanger sequencing of ADAM10 in 5 more patients from 4 unrelated Japanese families identified 4 different mutations in ADAM10. Recently POFUT1 gene was reported as one of the causative genes of DDD. Thus, we performed mutation search for POFUT1 in one patient who showed atypical skin manifestation as RAK. In this paper, we report that a POFUT1 mutation, but no ADAM10 mutation was detected in the patient, which makes further confirmation that RAK and DDD are genetically and clinicopathologically independent disorders distinct from each other. http://dx.doi.org/10.1016/j.jdermsci.2016.08.250