論文 Case of annular pustular psoriasis/circinate erythematous psoriasis induced by hydroxychloroquine in a patient with systemic lupus erythematosus: Possible association with CARD-14 mutation. The Journal of dermatology,48(9):e440-e442 2021(Sep.) Author:Hirotaka Midorikawa; Yukiko Kiniwa; Akane Minagawa; Kana Osawa; Takushi Shirai; Tasuku Sano; Kenta Nakamura; Atsuko Ashida; Ken-Ichi Ueno; Takuya Takeichi; Masashi Akiyama; Ryuhei Okuyama
Usefulness of monitoring circulating tumor cells as a therapeutic biomarker in melanoma with BRAF mutation. BMC cancer,21(1):287-287 2021(Mar. 17) Author:Yukiko Kiniwa; Kenta Nakamura; Asuka Mikoshiba; Atsuko Ashida; Yasuyuki Akiyama; Atsushi Morimoto; Ryuhei Okuyama Abstract:BACKGROUND: While molecularly targeted therapies and immune checkpoint inhibitors have improved the prognosis of advanced melanoma, biomarkers are required to monitor drug responses. Circulating tumor cells (CTCs) are released from primary and/or metastatic tumors into the peripheral blood. We examined whether CTCs have potential as biomarkers by checking the number of CTCs, as well as the BRAF genotype of individual CTCs, in melanoma patients undergoing BRAF/MEK inhibitor treatment. METHODS: CTCs were isolated from peripheral blood using a high-density dielectrophoretic microwell array, followed by labeling with melanoma-specific markers (MART-1 and/or gp100) and a leukocyte marker (CD45). The numbers of CTCs were analyzed in fifteen patients with stage 0-III melanoma. Furthermore, changes in CTC numbers were assessed in five patients with stage IV melanoma at four time points during BRAF/MEK inhibitor treatment, and the BRAF genotype was analyzed in CTCs isolated from one patient. RESULTS: We examined CTCs in patients with stage 0-III (five samples per stage: stage 0-I, stage II, and stage III), and detected CTCs even in patients with early disease (stage 0 and I). Interestingly, recurrence occurred in the lymph nodes of one stage I patient 2 years after the detection of a high number of CTCs in the patient's blood. The total number of CTCs in four of five patients with stage IV melanoma fluctuated in response to BRAF/MEK inhibitor treatment, suggesting that CTC number has potential for use as a drug response marker in advanced disease patients. Interestingly, one of those patients had CTCs harboring seven different BRAF genotypes, and the mutated CTCs disappeared upon BRAF/MEK inhibitor treatment, except for those harboring BRAFA598V. CONCLUSIONS: CTCs are present even in the early stage of melanoma, and the number of CTCs seems to reflect patients' responses to BRAF/MEK inhibitor treatment. Furthermore, genetic heterogeneity of BRAF may contribute to resistance to BRAF/MEK inhibitors. Our findings demonstrate the usefulness of CTC analysis for monitoring responses to targeted therapies in melanoma patients, and for understanding the mechanism of drug resistance.
Paraneoplastic pemphigus associated with Waldenström's macroglobulinemia. The Journal of dermatology,47(5):e200-e201 2020(May) Author:Takushi Shirai; Yukiko Kiniwa; Norito Ishii; Takashi Hashimoto; Yasushi Senoo; Kazuhisa Urushihata; Atsuko Ashida; Ryuhei Okuyama
Detecting copy number alterations of oncogenes in cell-free DNA to monitor treatment response in acral and mucosal melanoma. Journal of dermatological science,97(3):172-178 2020(Mar.) Author:Asuka Mikoshiba; Atsuko Ashida; Kaori Sakaizawa; Yukiko Kiniwa; Ryuhei Okuyama Abstract:BACKGROUND: Reliable biomarkers are necessary for assessment of treatment responses. Acral and mucosal melanomas are commonly associated with copy number (CN) alterations rather than specific point mutations, with CN alterations inKIT, CDK4, and CCND1 occurring frequently. Cell-free DNA is released to peripheral blood by both normal and tumor cells, and therefore contains the same genetic alterations present in the source tumor. OBJECTIVE: To investigate the usefulness of detecting CN alterations in oncogenes in cell-free DNA for monitoring treatment response in acral and mucosal melanomas. METHODS: We isolated cell-free DNA from peripheral blood and assessed the CN alterations in the cell-free DNA. Using droplet digital PCR, we examined CN alterations ofKIT, CDK4, and CCND1 in tumors from 37 melanoma patients (acral, n = 27; mucosal, n = 10) and peripheral blood from 24 melanoma patients (acral, n = 17; mucosal, n = 7). RESULTS: CN gain was detected in at least one of the genes examined in 62.9 % (17/27) of acral melanomas and 70 % (7/10) of mucosal melanomas. CN gains were also detected in the plasma of some patients. Furthermore, plasma CN ratio was correlated with clinical condition. This correlation was especially clear in patients with high CN ratios in tumors and high tumor burdens. CONCLUSION: Plasma CN ratios may be useful for evaluating treatment responses in patients with acral and mucosal melanoma.
第3選択としてダカルバジンを投与した悪性黒色腫3症例の臨床効果 Skin Cancer (Web),33(3) 2019 Author:中村謙太; 芦田敦子; 中村麗那; 木庭幸子; 奥山隆平
2.メラノーマの遺伝子異常 日本皮膚科学会雑誌,128(6) 2018 Author:芦田敦子
Circulating Tumour DNA for Monitoring Treatment Response to Anti-PD-1 Immunotherapy in Melanoma Patients. Acta dermato-venereologica,97(10):1212-1218 2017(Nov. 15) Author:Atsuko Ashida; Kaori Sakaizawa; Hisashi Uhara; Ryuhei Okuyama Abstract:Anti-programmed cell death-1 (anti-PD-1) antibody shows high therapeutic efficacy in patients with advanced melanoma. However, assessment of its therapeutic activity can be challenging because of tumour enlargement associated with intratumoural inflammation. Because circulating tumour DNA (ctDNA) correlates with tumour burden, we assessed the value of ctDNA levels as an indicator of tumour changes. Quantification of ctDNA (BRAFmutant or NRASmutant) levels by droplet digital PCR in 5 patients with BRAF or NRAS mutant melanoma during the treatment course showed dynamic changes corresponding to radiological and clinical alterations. In 3 cases in which the anti-PD-1 antibody was effective, ctDNA levels decreased within 2-4 weeks after treatment initiation. In 2 cases in which the anti-PD-1 antibody was ineffective, ctDNA levels did not decrease after treatment initiation. ctDNA could be a useful biomarker to predict early response to treatment in patients with advanced melanoma treated with anti-PD-1 immunotherapy.
Detection of BRAF(V600K) mutant tumor-derived DNA in the pleural effusion from a patient with metastatic melanoma CLINICAL CHEMISTRY AND LABORATORY MEDICINE,55(4):E92-E95 2017(Apr.) Author:Kaori Sakaizawa; Atsuko Ashida; Hisashi Uhara; Ryuhei Okuyama
Detection of BRAFV600K mutant tumor-derived DNA in the pleural effusion from a patient with metastatic melanoma Clin Chemi Lab Med,55(4):e92-e95 2017(Mar.) Author:Kaori Sakaizawa, Ashida A, Uhara H , Okuyama R
Distinct phenotype of epidermolysis bullosa simplex with infantile migratory circinate erythema due to frameshift mutations in the V2 domain of KRT5 European Academy of Dermatology and VenereologyActa Derm Venereol ,31(5):e241-e243 2016(Nov.) Author:Y.Kumagai, N. Umegaki-Arao, T. Sasaki,Y. Nakamura, H. Takahashi, A. Ashida, Y. Tsunemi,M. Kawashima, A. Shimizu, A. Ishiko, K. Nakamura,H. Tsuchihashi, M. Amagai, A. Kubo
Quantitative analysis of the BRAF V600E mutation in circulating tumor-derived DNA in melanoma patients using competitive allele-specific TaqMan PCR. International journal of clinical oncology,21(5):981-988 2016(Oct.) Author:Atsuko Ashida; Kaori Sakaizawa; Asuka Mikoshiba; Hisashi Uhara; Ryuhei Okuyama Abstract:BACKGROUND: BRAF V600E is a common mutation in melanoma, and BRAF inhibitors are effective in treating of BRAF mutation-positive melanoma. DNA carrying this mutation is released from melanoma cells into the circulation. As such, circulating tumor-derived DNA (ctDNA) in peripheral blood represents a novel biomarker for evaluating tumor features in cancer patients. However, ctDNA is present in the peripheral blood at very low levels, which makes the detection of specific mutations in this DNA a challenge. Competitive allele-specific TaqMan PCR (castPCR), a straightforward commercially available assay, is a sensitive technique for quantitating a small amount of DNA. METHODS: The level of BRAF V600E ctDNA was quantified by castPCR in 26 consecutive plasma samples from six melanoma patients. RESULTS: The castPCR assay was performed using a mixture of BRAF V600E DNA and BRAF wild DNA and found to be able to detect BRAF V600E at a fractional abundance of ≥0.5 % in 2- to 10-ng samples of genomic DNA. Cell-free DNA was then extracted from peripheral blood samples collected from six patients with melanoma harboring the BRAF V600E mutation. BRAF V600E ctDNA was detected in three patients, at a fractional abundance of between 1.28 and 58.0 % of total BRAF cell-free DNA. The abundance of BRAF V600E ctDNA correlated with tumor burden, as determined by computed tomography imaging. In two cases, an increase in the level of BRAF V600E ctDNA preceded exacerbation of clinical symptoms. CONCLUSION: The castPCR assay can detect and quantitate small amounts of BRAF V600E ctDNA in samples containing large amounts of BRAF wild cell-free DNA. Thus, we suggest that the castPCR assay is suitable for monitoring ctDNA in the plasma of melanoma patients.
Acquired epidermodysplasia verruciformis associated with human papillomavirus type 47 in a HIV-infected patient. The Journal of dermatology,43(6):717-8 2016(Jun.) Author:Atsuko Ashida; Akira Shimizu; Ryuhei Okuyama
Quantitative analysis of the BRAFV600E mutation in circulating tumor-derived DNA using competitive allele-specific TaqMan PCR in melanoma patients Int J Clin Oncol.,21(5):981-988 2016(May) Author:Ashida A, Sakaizawa K, Mikoshiba A, Uhara H , Okuyama R
Inhibition of epidermal growth factor receptor induces tumor necrosis factor-α via activation of peroxisome proliferator-activated receptor-γ and nuclear factor-κB in sebocytes: A possible pathogenesis of papulopustular rash. Journal of dermatological science,82(1):53-6 2016(Apr.) Author:Atsuko Ashida; Eisaku Ogawa; Hisashi Uhara; Tomonobu Koizumi; Ryuhei Okuyama
Melanoma with BRAF Mutation in Circulating Cell-free DNA despite no Mutation in the Primary Lesion: A Case Report. Acta dermato-venereologica,96(1):128-9 2016(Jan.) Author:Atsuko Ashida; Hisashi Uhara; Asuka Mikoshiba; Kaori Sakaizawa; Naomi Kumagai; Hiroshi Koga; Ryuhei Okuyama
Histopathological improvement of scleroderma induced by paclitaxel in a patient with breast cancer. The Journal of dermatology,42(12):1198-9 2015(Dec.) Author:Atsuko Ohashi; Akane Minagawa; Atsuko Ashida; Hiroshi Koga; Hisashi Uhara; Ryuhei Okuyama
Clinical characteristics associated with BRAF, NRAS and KIT mutations in Japanese melanoma patients. Journal of dermatological science,80(1):33-7 2015(Oct.) Author:Kaori Sakaizawa; Atsuko Ashida; Aya Uchiyama; Takamichi Ito; Yasuhiro Fujisawa; Dai Ogata; Shigeto Matsushita; Kazuyasu Fujii; Satoshi Fukushima; Yoshitsugu Shibayama; Naohito Hatta; Tatsuya Takenouchi; Jiro Uehara; Ryuhei Okuyama; Naoya Yamazaki; Hisashi Uhara Abstract:BACKGROUND: The importance of the genetic background of melanoma cells to the individual susceptibility to treatment has become apparent. In Caucasians, BRAF mutations are frequently detected in lesions on the skin of younger patients compared to NRAS and KIT mutations. However, clinical and pathological characteristics associated with BRAF, NRAS and KIT mutations have not been fully evaluated in East Asians. OBJECTIVE: To clarify clinical and pathological characteristics associated with BRAF, NRAS and KIT mutations in Japanese melanoma patients. METHODS: Clinical data were retrospectively collected from 11 hospitals in Japan. BRAF, NRAS and KIT mutations were evaluated with polymerase chain reaction and Sanger sequencing. The relationships between these gene mutations and pathological and clinical findings were analyzed. RESULTS: The number of cases examined was 171 (primary: 135, metastases: 11, paired: 25), and all were Japanese patients. The detection rates of BRAF, NRAS and KIT mutations were 30.4%, 12.3% and 12.9%, respectively. Compared with the wild type, the presence of BRAF mutations was significantly associated with younger age (median, 50.0 years vs. 70.0 years, p<0.001). BRAF mutation was frequently detected in the lesions of the scalp (80%; 4/5), trunk (72.0%; 18/25), extremities (56.7%; 17/30) and neck (44.4%; 4/9), and the least prevalent were the face (22.2%; 2/9), nail (12.5%; 3/24), palm or sole (8.9%; 4/45) and mucosa (0%). NRAS mutations were prevalent in the face (33.3%) and palm or sole (20.0%), and the median age of these patients was 70.5 years. A KIT mutation was observed in the nail apparatus (25%), palm or sole (15.6%) and mucosa (18.2%). The median age of the patients with a KIT mutation was 63.0 years. Heterogeneity of mutations between primary and metastatic lesions was detected in six of 25 cases (24%). Solar elastosis was identified in 12 of 71 cases (15.3%), among which four cases harbored BRAF(V600E) (2 cases), BRAF(V600K), NRAS(Q61K) or NRAS(Q61L), respectively. CONCLUSION: Some clinical characteristics associated with BRAF, NRAS and KIT mutations were observed in Japanese patients, and we observed both similarities to and differences from those of Caucasians. Our findings could provide useful information in efforts to clarify the tumor genesis of malignant melanomas.
An Initial Case Report of a Laparoscopic Spleen-Preserving Distal Pancreatectomy for a Patient with a Pancreatic Metastasis of Malignant Melanoma The Shinshu medical journal,63(1):19-24 2015(Jan.) Author:Motoyama H, Kobayashi A, Yokoyama T, Shimizu A, Furusawa N , Sakai H, Kitagawa N, Ashida A, Okuyama R, Miyagawa S
Toll-like receptor 4 signaling promotes the migration of human melanoma cells. The Tohoku journal of experimental medicine,234(1):57-65 2014(Sep.) Author:Yuko Takazawa; Yukiko Kiniwa; Eisaku Ogawa; Aya Uchiyama; Atsuko Ashida; Hisashi Uhara; Yasufumi Goto; Ryuhei Okuyama Abstract:Immune cell Toll-like receptors (TLRs) recognize conserved microbial components, leading to immune and inflammatory responses. However, TLRs are also expressed in cancer cells, including melanoma cells, which express TLR2-4. TLR4 ligands have received attention as immunotherapies; therefore, we assessed the expression of TLR4 in human melanoma specimens (29 primary lesions and 28 metastatic lesions) representing different types of melanoma. A high percentage (≥ 90%) of melanoma lesions expressed TLR4, as judged by immunohistochemistry. Next, the role of TLR4 in cell proliferation and migration was assessed using the TLR4-positive (TLR4(+)) melanoma cell lines 501mel and 888mel, and TLR4-negative (TLR4(‒)) 928mel melanoma cells. Lipopolysaccharide (LPS), a TLR4 agonist, increased the proliferation of TLR4(+) melanoma cells but not of TLR4(‒) 928mel cells. The proliferation-inducing effect of LPS in 888mel cells was abolished by blockade of TLR4 signaling via treatment with short interfering RNA (siRNA) targeting TLR4 or myeloid differentiation primary response gene 88 (MyD88), a molecule downstream of TLR4. However, knockdown of TLR4 or MyD88 expression did not affect the LPS-induced proliferation of 501mel cells, suggesting that residual TLR4 signaling is sufficient to maintain cell proliferation. By contrast, LPS increased the migration of TLR4(+) melanoma cells, and this effect was substantially inhibited by TLR4 or MyD88 knockdown. Furthermore, TLR4 knockdown decreased cell migration even in the absence of LPS, suggesting the presence of an endogenous TLR4 ligand(s) in melanoma cells. TLR4 signaling may contribute to melanoma progression, and caution should be exercised when using TLR4 ligands as adjuvant therapy for cancer.
5-Hydroxymethylcytosine as a useful marker to differentiate between malignant melanomas and benign melanocytic nevi. Journal of dermatological science,73(2):161-3 2014(Feb.) Author:Ryuhei Uchiyama; Hisashi Uhara; Aya Uchiyama; Eisaku Ogawa; Yuko Takazawa; Atsuko Ashida; Hiroshi Koga; Koichi Hayashi; Yukiko Kiniwa; Ryuhei Okuyama
Successful treatment of rheumatoid vasculitis-associated skin ulcer with a TNF-α antagonist. International journal of dermatology,53(2):e154-6 2014(Feb.) Author:Atsuko Ashida; Hiroshi Murata; Yasutomo Mikoshiba; Atsuko Ohashi; Aya Kobayashi; Hiroshi Koga; Hisashi Uhara; Ryuhei Okuyama
Fibrosarcomatous variant of dermatofibrosarcoma protuberans with pancreatic metastasis. International journal of dermatology,53(2):e140-2 2014(Feb.) Author:Hiroshi Murata; Atsuko Ohashi; Atsuko Ashida; Hisashi Uhara; Ryuhei Okuyama; Takenari Nakata; Kunihiko Shingu
Coexpression of EpCAM, CD44 variant isoforms and claudin-7 in anaplastic thyroid carcinoma. PloS one,9(4):e94487 2014 Author:Toshihiro Okada; Teruo Nakamura; Takayuki Watanabe; Naoyoshi Onoda; Atsuko Ashida; Ryuhei Okuyama; Ken-ichi Ito Abstract:BACKGROUND: Anaplastic thyroid cancer is considered to be one of the most aggressive human malignancies, and the mean survival time after diagnosis is approximately six months, regardless of treatments. This study aimed to examine how EpCAM and its related molecules are involved in the characteristics of anaplastic thyroid carcinoma. METHODOLOGY/PRINCIPAL FINDINGS: Two differentiated thyroid cancer cell lines (TPC-1 and FTC-133), and two anaplastic thyroid cancer cell lines (FRO, ACT-1) were analyzed for expression of CD44 standard isoform (CD44s), CD44 variant isoforms, and EpCAM, and human aldehyde dehydrogenase-1 (ALDH1) enzymatic activity using flow cytometry. CD44s expression was higher in TPC-1 and FTC-133 than in the FRO and ACT-1, whereas ALDH1 activities were higher in FRO and ACT-1 than in TPC-1 and FTC-133. An inverse correlation between CD44s expression and ALDH1 activity was observed in all thyroid cancer cell lines. As for the expressions of CD44 variant isoforms, ACT-1 showed higher and FRO showed moderate CD44v6 expressions, whereas either TPC-1 or FTC-133 showed negative CD44v6 expression. EpCAM expressions in FRO and ACT-1 were higher than those in TPC-1 and FTC-133, and EpCAM expressions inversely correlated with those of CD44s. A positive correlation was observed between EpCAM expression and ALDH1 activity in thyroid cancer cell lines. In the RT-PCR analysis, the expression levels of EpCAM, caludin-7 and ALDH1 in FRO and ATC-1 cells were significantly higher than those in TPC-1 and FTC-133 cells. In clinical specimens of thyroid cancers, nuclear expression of EpCAM and high expression of CD44v6 were detected significantly more frequently in anaplastic carcinomas. CONCLUSIONS/SIGNIFICANCE: Our study suggests the possibility that EpCAM, together with CD44v6 and claudin-7 as well as ALDH1, may be involved in the development of the aggressive phenotype of anaplastic thyroid carcinoma. Our findings may suggest a novel therapeutic strategy for treatment of anaplastic thyroid carcinoma.
NRAS mutations in primary and metastatic melanomas of Japanese patients. International journal of clinical oncology,19(3):544-8 2014 Author:Hisashi Uhara; Atsuko Ashida; Hiroshi Koga; Eisaku Ogawa; Aya Uchiyama; Ryuhei Uchiyama; Koichi Hayashi; Yukiko Kiniwa; Ryuhei Okuyama Abstract:BACKGROUND: Characterization of the MAPK signaling pathway in melanoma has led to the development of MEK inhibitors for the treatment of NRAS-mutated melanoma. The success of molecular-targeted therapies underscores the need to identify mutations in target genes. Most of the current data on genetic mutations have been obtained from Caucasian melanoma patients, and screenings of Asian populations are limited. OBJECTIVE: The aim of the present study was to examine NRAS mutations in primary and metastatic lesions of Japanese melanoma patients. METHODS: Clinical melanoma specimens were collected from 127 Japanese patients, including primary (n = 67), metastatic (n = 25) and paired primary and metastatic lesions (n = 35). NRAS mutations in exons 1 and 2 were assessed by polymerase chain reaction and Sanger sequencing. RESULTS: The incidence of NRAS mutations was 7.1 %. NRAS (Q61) was the predominant genetic alteration (77.8 %). NRAS mutations were most frequently detected in acral melanomas (9.3 %), followed by melanomas without chronic sun-induced damage (7.0 %) and mucosal melanomas (4.8 %), and were not detected in melanomas with chronic sun-induced damage. In addition, NRAS mutations were more prevalent in the extremities than in other sites. The NRAS sequence in metastatic lesions did not match that of the primary tumor in one case. CONCLUSION: The frequency of NRAS mutations is lower in the Asian population than in Caucasian patients. The observed heterogeneity of melanoma suggests that genotyping of both primary and metastatic lesions is important to identify candidate patients for molecular-targeted therapies.
Combination chemotherapy of carboplatin and paclitaxel for metastatic melanoma. The Journal of dermatology,40(12):1050-1 2013(Dec.) Author:Yuki Sato; Hisashi Uhara; Atsuko Ashida; Ryuhei Okuyama
A case of hypocomplementaemic urticarial vasculitis with a high serum level of rheumatoid factor. The Australasian journal of dermatology,54(3):e62-3 2013(Aug.) Author:Atsuko Ashida; Hiroshi Murata; Atsuko Ohashi; Eisaku Ogawa; Hisashi Uhara; Ryuhei Okuyama Abstract:We report a case of hypocomplementaemic urticarial vasculitis with an elevated serum rheumatoid factor level. Hypocomplementaemic urticarial vasculitis is an immune complex-mediated disease characterised by urticarial eruptions. High levels of rheumatoid factor may be associated with hypocomplementaemia due to the consumption of complement, because the rheumatoid factor can form immune complexes with immunoglobulin. It is necessary to pay attention to the amounts of complement in cases of urticarial eruptions with elevated rheumatoid factor level. The eruptions were relieved with a combination of prednisolone and colchicine.
Low p53 positivity in verrucous skin lesion in diabetic neuropathy occurring on the dorsum of the foot International Journal of Dermatology,52(3):378-380 2013(Mar.) Author:Atsuko Ashida; Yukiko Kiniwa; Aya Kobayashi; Kazuhiko Matsumoto; Ryuhei Okuyama
Low p53 positivity in verrucous skin lesion in diabetic neuropathy occurring on the dorsum of the foot Int J Dermatol,52:378-380 2013 Author:Ashida A, Kiniwa Y, Kobayashi A, Matsumoto K, Okuyama R
Clinical courses of drug-induced hypersensitivity syndrome treated without systemic corticosteroids. J Eur Acad Derm Venereol,27:722-726 2013 Author:Uhara H, Saiki M, Kawachi S, Ashida A, Oguchi S, Okuyama R
Case of a cutaneous angiomyolipoma in the ear JOURNAL OF DERMATOLOGY,39(9):808-809 2012(Sep.) Author:Yasutomo Mikoshiba; Hiroshi Murata; Atsuko Ashida; Nana Saito; Hiroshi Koga; Hisashi Uhara; Ryuhei Okuyama
Assessment of BRAF and KIT mutations in Japanese melanoma patients JOURNAL OF DERMATOLOGICAL SCIENCE,66(3):240-242 2012(Jun.) Author:Atsuko Ashida; Hisashi Uhara; Yukiko Kiniwa; Misae Oguchi; Hiroshi Murata; Yasufumi Goto; Aya Uchiyama; Eisaku Ogawa; Koichi Hayashi; Hiroshi Koga; Ryuhei Okuyama
Assessment of BRAF and KIT mutations in Japanese melanoma J Dermatological Science,66(3):240-242 2012(Jun.) Author:Ashida A, Uhara H, Kiniwa Y, Oguchi M, Murata H, Goto Y, Uchiyama A, Ogawa E, Hayashi K, Koga H, Okuyama R
A case of epidermolysis bullosa acquisita associated with laryngeal stenosis. Acta dermato-venereologica,92(1):93-4 2012(Jan.) Author:Yukiko Kiniwa; Atsuko Ashida; Atsuko Ohashi; Ryosuke Kitoh; Shunpei Fukuda; Takashi Hashimoto; Ryuhei Okuyama
Case of a cutaneous angiomyolipoma in the ear J Dermatol,39:808-809 2012 Author:Mikoshiba Y, Murata H, Ashida A, Saito N, Koga H, Uhara H, Okuyama R
Assessment of BRAF and KIT mutations in Japanese melanoma patients J Dermatol Sci,66::240-242 2012 Author:Ashida A, Uhara H, Kiniwa Y, Oguchi M, Murata H, Goto Y, Uchiyama A, Ogawa E, Hayashi K, Koga H, Okuyama R
Thermal angioedema induced by hot water Acta Derm Venereol,91:343-344 2011 Author:Kobayashi A, Uhara H, Ashida A, Kiniwa Y, Okuyama R
NADPH oxidase 4 contributes to transformation phenotype of melanoma cells by regulating G2-M cell cycle progression. Cancer research,69(6):2647-54 2009(Mar. 15) Author:Maki Yamaura; Junji Mitsushita; Shuichi Furuta; Yukiko Kiniwa; Atsuko Ashida; Yasuhumi Goto; Wei H Shang; Makoto Kubodera; Masayoshi Kato; Minoru Takata; Toshiaki Saida; Tohru Kamata Abstract:Generation of reactive oxygen species (ROS) has been implicated in carcinogenic development of melanoma, but the underlying molecular mechanism has not been fully elucidated. We studied the expression and function of the superoxide-generating NADPH oxidase (Nox)4 in human melanoma cells. Nox4 was up-regulated in 13 of 20 melanoma cell lines tested. Silencing of Nox4 expression in melanoma MM-BP cells by small interfering RNAs decreased ROS production and thereby inhibited anchorage-independent cell growth and tumorigenecity in nude mice. Consistently, a general Nox inhibitor, diphenylene iodonium, and antioxidants vitamine E and pyrrolidine dithiocarbamate blocked cell proliferation of MM-BP cells. Flow cytometric analysis indicated that Nox4 small interfering RNAs and diphenylene iodonium induced G(2)-M cell cycle arrest, which was also observed with another melanoma cell line, 928mel. This was accompanied by induction of the Tyr-15 phosphorylated, inactive form of cyclin-dependent kinase 1 (a hallmark of G(2)-M checkpoint) and hyperphosphorylation of cdc25c leading to its increased binding to 14-3-3 proteins. Ectopic expression of catalase, a scavenger of ROS, also caused accumulation of cells in G(2)-M phase. Immunohistochemistry revealed that expression of Nox4 was detected in 31.0% of 13 melanoma patients samples, suggesting the association of Nox4 expression with some steps of melanoma development. The findings suggest that Nox4-generated ROS are required for transformation phenotype of melanoma cells and contribute to melanoma growth through regulation of G(2)-M cell cycle progression.
Pathological activation of KIT in metastatic tumors of acral and mucosal melanomas. International journal of cancer,124(4):862-8 2009(Feb. 15) Author:Atsuko Ashida; Minoru Takata; Hiroshi Murata; Kenji Kido; Toshiaki Saida Abstract:Recent studies showed KIT gene aberrations in a substantial number of melanomas on acral skin and mucosa, suggesting the therapeutic benefit of tyrosine kinase inhibitors, such as imatinib. We therefore examined the expression and mutations of KIT in 4 primary and 24 metastatic acral and mucosal melanomas. Immunohistochemistry revealed moderate or strong KIT protein expression in 13 (48%) tumors. Sequence analysis revealed K642E and D820Y mutations in two metastases. Amplification of KIT was identified by real-time PCR in 4 tumors, including one that had K642E. Western blot analysis showed phosphorylation of the KIT receptor in 8 (62%) of 13 cryopreserved samples, indicating the frequent pathological activation of the receptor in vivo. Phosphorylation of KIT protein was detected in 2 tumors harboring KIT mutations, as well as in one tumor with KIT gene amplification. Furthermore, 5 tumors without detectable KIT gene aberrations showed phosphorylation of the KIT receptor. Expression of stem cell factor (SCF) in melanoma cells as well as stromal cells suggests SCF/KIT autocrine and paracrine activation in these tumors. Finally, we found significant growth suppressive effects of sunitinib in two acral melanoma cell lines; one harboring the D820Y mutation and one showing SCF-dependent KIT activation. These results show pathological activation of KIT in a substantial number of metastatic tumors of acral and mucosal melanomas, and suggest a potential therapeutic benefit of sunitinib for these melanomas.
Interferon-beta therapy for malignant melanoma: the dose is crucial for inhibition of proliferation and induction of apoptosis of melanoma cells. Archives of dermatological research,300(6):297-301 2008(Jul.) Author:Hitomi Kubo; Atsuko Ashida; Kazuhiko Matsumoto; Toshiro Kageshita; Akifumi Yamamoto; Toshiaki Saida Abstract:We investigated the anti-tumor effect of human interferon-beta (HuIFN-beta) against malignant melanoma. In vitro study revealed that HuIFN-beta not only inhibited proliferation of melanoma cells (seven cell lines: MM-AN, MM-BP, MM-LH, MM-RU, PM-WK, RPM-EP, RPM-MC) but also induced apoptosis in a dose dependent fashion, though the sensitivity to HuIFN-beta was different among cell lines. In addition, we administered HuIFN-beta into cutaneous metastatic lesions of melanoma and evaluated clinical and histopathological effects. Although the size of the metastatic cutaneous lesion did not change by the intralesional injection of HuIFN-beta, histopathological examination revealed apoptotic changes of melanoma cells along with dense lymphohistiocytic infiltration. The present study confirmed direct and indirect inhibitory effects of HuIFN-beta on human melanoma cells and suggests that local higher concentration of HuIFN-beta is needed to eradicate melanoma lesions.
Establishment of a novel melanoma cell line SMYM-PRGP showing cytogenetic and biological characteristics of the radial growth phase of acral melanomas. Cancer science,98(7):958-63 2007(Jul.) Author:Hiroshi Murata; Atsuko Ashida; Minoru Takata; Maki Yamaura; Boris C Bastian; Toshiaki Saida Abstract:We established a novel melanoma cell line, SMYM-PRGP, which was non-tumorigenic in vivo, from an acral melanoma in radial growth phase under a low-oxygen environment. SMYM-PRGP was wild-type for known mutation sites in the BRAF and NRAS genes, and showed focal amplification of the human telomerase reverse transcriptase and cyclin D1 genes as well as the fibroblast growth factor-3 and fibroblast growth factor-4 genes. Neither mutation nor copy number loss of the CDKN2A gene was observed. The p16(INK4A) protein was expressed at a level equal to that in normal melanocytes. Among the various melanocyte growth factors added to the culture of SMYM-PRGP cells, endothelin-1 was the strongest growth stimulator, the effect of which was significantly augmented by the addition of calcium chloride. The growth stimulatory effect of endothelin-1 was shown to be mediated via the endothelin B receptor. The protein level of cyclin D1 in SMYM-PRGP cells was approximately 10 times higher than that in normal melanocytes. Although the stimulation with endothelin-1 plus calcium chloride increased cyclin D1 protein levels after 4-6 h, the level of phosphorylated retinoblastoma protein did not increase, suggesting that overexpression of cyclin D1 protein may have little effect on cell cycle progression but rather act as a pro-survival factor. SMYM-PRGP is an excellent tool for investigating the development and progression of acral melanoma.
Quantitative analysis of the BRAFV600E in circulating tumor-drived DNA using competitive allele-specific TaqMan PCR in melanoma patients 30回表皮細胞研究会≪弘前） 2016(Nov.) Presenter:芦田敦子、境澤香里、宇原久、奥山隆平
Evaluation of response to nivolumab treatment in patients with metastatic melanoma using circulating tumor DNA 75回日本癌学会学術総会（横浜） 2016(Oct.) Presenter:芦田敦子、境澤香里、宇原久、奥山隆平
Epidermal growth factor receptor inhibitor induces tumor necrosis factor-α via activation of the PPARγ and NF-κB from sebocytes 40回日本研究皮膚科学会学術大会総会（岡山） 2015(Dec.) Presenter:Atsuko Ashida, Eisaku Ogawa, Ryuhei Uchiyama, Hisashi Uhara, Ryuhei Okuyama
Detection of BRAFV600Ecirculating tumor DNA using castPCR in patients with advanced melanoma 74回日本癌学会学術総会（名古屋） 2015(Oct.) Presenter:Atsuko Ashida, Kaori Sakaizawa, Hisashi Uhara, Ryuhei Okuyama
NRAS mutation in Japanese melanoma patients 8th World Congress of Melanoma（ハンブルグ） 2013(Jul.) Presenter:Hisashi Uhara, Atsuko Ashida, Aya Uchiyama, Richie Okuyama
Clinical and genetic characteristics of cutaneous melanoma with clinical history of early childhood onset JOURNAL OF INVESTIGATIVE DERMATOLOGY,137(10):S196-S196 2017(Oct.) Author:A. Minagawa; H. Uhara; K. Sakaizawa; H. Koga; A. Ashida; R. Okuyama
NADPH oxidase(Nox)4によるG2-M細胞周期進行の新しい調節機構(A Novel link of NADPH oxidase(Nox)4 redox-signaling to G2-M cell cycle progression in melanoma cell proliferation) 日本生化学会大会プログラム・講演要旨集,82回:3T10a-5 2009(Sep.) Author:山浦 真貴; 満下 淳地; 古田 秀一; 木庭 幸子; 芦田 敦子; 後藤 泰文; 加藤 真良; 高田 実; 斎田 敏明; 鎌田 徹
メラノーマでは、RAS/RAF/MEK/ERKシグナルはcyclin D、cyclin Eの双方を制御する(Both cyclin D and cyclin E are downstream targets of the RAS/RAF/MEK/ERK pathway in melanoma cells) 日本癌学会総会記事,68回:485-486 2009(Aug.) Author:村田 浩; 芦田 敦子; 後藤 康文; 木藤 健治; 高田 実
転移性肢端型・粘膜型黒色腫では野生型KITが活性化している(Pathological activation of wild-type KIT is common in metastatic tumors of acral and mucosal melanomas) 日本癌学会総会記事,67回:222-222 2008(Sep.) Author:芦田 敦子; 村田 浩; 木藤 健治; 高田 実; 斎田 俊明
水平増殖期末端黒子型悪性黒色腫細胞株の樹立とその性質の解析(Establishment and characterization of a cell line derived from the radial growth phase acral melanoma) 日本癌学会総会記事,64回:206-206 2005(Sep.) Author:村田 浩; 芦田 敦子; 高田 実; 斎田 俊明
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