DNA repair synthesis following irradiation with 254-nm and 312-nm ultraviolet light is not diminished in fibroblasts from patients with dysplastic nevus syndrome

The DNA excision repair capacity of 23 primary fibroblast lines from patients with dysplastic nevus syndrome was investigated and DNA repair synthesis (unscheduled DNA synthesis) was determined after UV exposure. Seventeen fibroblast lines from normal donors served as controls. The dose/response experiments included up to ten dose levels and two wavelength ranges: UV-C (using a low-pressure mercury lamp emitting predominantly 254-nm light) and UV-B (artificial sunlamp radiation centering around 312-nm light). For each dose level, silver grains over fibroblast nuclei were counted by visual inspection. Twelve cell lines were also evaluated for both UV wavelength ranges using a new semi-automatic image analyzing system. This system included components for rapid sequential identification of both fibroblast nuclei and silver grains sited above them. Silver grains over 100 nuclei were determined for each UV dose level. Dose/response curves were established and analyzed by linear regression. As a quantitative term for assessing DNA excision repair capacity of a cell line we calculated the linear increase (G ₀) in the number of grains per nucleus, when the UV dose was multiplied by the factor e (i.e. 2.72). The sensitivity of grain detection and resolution ofoverlapping grains was approximately threefold better in visual than in automatic counting, especially when there were more than 70 grains over nuclei. The time recired for visual conting, however, was tenfold that of automatic counting. The varianceweighted meanG ₀ ^v,w of all fibroblast lines from patients with dysplastic nevus syndrome was found to be 79.1 (±1.8-grains/nucleus, that of fibroblast lines from normal donors was 74.2 (±1.7) grains/nucleus. This difference revealed a slightly better repair capability for cell lines from patients but was at the borderline of detection and, therefore, should not be overinterpreted. From the experimental accuracy achieved by determination of the varianceweighted means of the two groups, we would have been able to detect a difference of 7 and more grains [> 2 x (_normal + _patients)]. The variance-weighted meanG _o ^v,w of all fibroblast lines from patients with dysplastic nevus syndrome was found to be 76.4 (±1.4) grains/nucleus, whereas that of fibroblast lines from normal donors was only 66.6 (±1.8) grains/nucleus. This difference was statistically significant and, contrary to expectation, revealed better, not worse post-UV DNA repair capability in cell lines from patients that in those from normal donors. From the experimental accuracy achieved by determination of the variance-weighted means of the two groups, we would have been able to detect a difference of 6.4 or more grains [> 2 x (_normal + _patients)]. Variation between cell lines belonging to the same group was expressed by the standard deviation. On average, the standard deviation was in the range 18.2–21.1 grains/nucleus. This variation did not reflect experimental inaccuracy but different responses of individual cell lines to UV irradiation. On the basis of our data, we consider the hypothesis that patients with dysplastic nevus syndrome are prone to melanoma development because of a general defect in post-UV DNA repair to be improbable.This work was supported by the Deutsche Forschungsgemeinschaft, SFB 136     

Evaluation of merbarone (NSC 336628) in disseminated malignant melanoma

Merbarone, NSC 336628, is an investigational anticancer drug with activity against experimental animal tumors including melanoma. This paper presents results of a Phase II clinical study of merbarone in patients with biopsy proven stage IV malignant melanoma without prior chemotherapy and with no evidence of CNS involvement. Thirty-five patients with median age 58 (range 27–81), with performance status 0–2 were treated with merbarone 1000 mg/m²/day for five days by intravenous continuous infusion repeated every 3 weeks. All patients (21 males and 14 females) were evaluable for toxicity. Two patients were not evaluable for response having been removed from protocol treatment due to toxicity and received other treatment during the first course of chemotherapy. Among the evaluable patients there was one complete response in a supraclavicular lymph node lasting four months and one partial liver response lasting three months. The remaining thirty-one patients were non-reponders. Of these one had a stable disease lasting 21 months. The overall objective response rate was 6% (2/35) with a 95% confidence interval of 1%–19%. Twenty-six of the 35 patients have died. The estimated median survival of the entire group was 9 months with a 95% confidence interval of six to eleven months. Renal toxicity was dose-limiting and manifested as increasing serum creatinine (54% of patients), proteinuria (51%) and hematuria (9%). One patient experienced grade 4 creatinine increase, proteinuria and acute renal failure. Other toxicities included nausea (71%), vomiting (51%), malaise (23%), weakness (20%), alopecia (17%), diarrhea (17%), anorexia (14%), transaminase (SGOT, SGPT) increase (14%), constipation (14%), alkaline phosphatase or 5nucleotidase increase (9%), and fever (9%). Hematologictoxicity (granulocytopenia, leukopenia, and anemia) was generally mild and infrequent (29%, only one patient had grade 4 granulocytopenia). Overall 9 patients (26%) had at least one grade 3 toxicity. We conclude that merbarone at this dose and schedule has detectable but minimal activity in the treatment of metastatic malignant melanoma and given the significant renal toxicity this schedule does not merit further evaluation in this disease.     

恶性黑素瘤患者外周血酪氨酸酶与临床分期的相关性研究

使用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）检测恶性黑素瘤患者外周血中酪氨酸酶ｍＲＮＡ,并与健康供血者和其它皮肤恶性肿瘤患者对照。结果：６８例恶性黑素瘤患者,临床Ⅰ期和Ⅱ期酪氨酸酶ｍＲＮＡ的ＲＴ－ＰＣＲ阳性率分别为６．５％和１３．３％,Ⅲ期为４２．９％,Ⅳ期（即有远距离转移者）为８７．５％（Ｐ＜０．００１）。说明外周血中酪氨酸酶ｍＲＮＡ阳性率与临床分期有明显的关系,而与性别、肿瘤发病部位和病理学分类无关（Ｐ＞０．０５）。在健康供血者和其他皮肤恶性肿瘤患者血液中酪氨酸酶ｍＲＮＡ的ＲＴ－ＰＣＲ均为阴性。提示酪氨酸酶ｍＲＮＡ具有特异性。     

小鼠黑色素瘤凝集素的制备和特性分析

探讨Ｓ型凝集素生化特性及作用。方法：利用亲合层析及凝胶过滤层析从小鼠黑色素瘤中提 取小鼠黑色素瘤凝集素（ＭＭＡ）。结果：ＭＭＡ的亚基分子量为１２．４ ｋＤ,以同型二聚体存在,特异性识别含多聚乙 酰氨基乳糖的糖链,活性依赖于巯基而非钙离子。结论：ＭＭＡ为Ｓ型凝集素。     

Primary anorectal malignant melanoma: Report of a case

(Received for publication on Aug. 18, 1997; accepted on May 15, 1998)     

A phase II study of Didemnin B (NSC 325319) in advanced malignant melanoma: an Eastern Cooperative Oncology Group study (PB687)

Purpose: Didemnin B is a novel marine natural product cyclic depsipeptide containing unusual amino acid moieties. This agent demonstrates promising preclinical antitumor activity, including activity against B16 melanoma and against melanoma isolates in the human tumor stem cell assay.Methods: We conducted a phase II study of Didemnin B, given in Cremophor, at a starting dose of 4.2 mg/m2/IV q 28 days. Patients with measurable metastatic or advanced malignant melanoma were eligible. All patients were previously untreated with chemotherapy and had performance status 0 or 1. Doses were escalated to 4.9 and 5.6 mg/m2 in cycles 2 and 3, respectively.Results: Nineteen patients were entered and treated with a median of one cycle per patient. Eight of these patients went off study for toxicity including 7 with anaphylactoid reactions in the first or second cycle. One patient went off study after 3 cycles with severe myopathy, a newly described toxicity. Two were not evaluable for response and five were considered stable, including one patient with a transient PR of soft tissue disease in the first cycle. Another patient had stable disease for twelve cycles before progressing and one went off study electively after 3 cycles, for a total of 7 patients with stable disease. One patient with a measurable partial remission (PR) and went off study after three cycles due to severe myopathy, a then newly-described toxicity. No hematologic toxicity was seen. Nausea and vomiting were controlled with anti-emetics.Conclusions: This study was indeterminate with respect to the activity of Didemnin B in melanoma. Signs of activity were seen, particularly in soft tissue masses, though a large number of patients could not be evaluated fully for activity due to the occurrence of anaphylactoid reactions. This study does not preclude a clinically important level of activity for Didemnin B.     

Neuroendocrine Tumors of the Head and Neck: A Selected Review with Emphasis on Terminology

Virtually every variant of neuroendocrine neoplasia can occur, at least rarely, in the head and neck region. This review focuses on the terminology surrounding neuroendocrine carcinomas of the larynx and their distinction from morphologically similar but biologically distinctive neoplasmas. It is suggested that rare typical laryngeal carcinoids be labeled as such. There is little evidence that these lesions are part of a morphologic continuum. In contrast, more common “carcinoid-like” carcinomas, previously referred to as “atypical carcinoids” are more appropriately labeled as “moderately differentiated neuroendocrine carcinomas”. These neoplasms should, in turn, be distinguished from “small cell neuroendocrine carcinomas,” although these latter two neoplasms do represent a morphologic and behavioral spectrum. Light microscopic and immunohistochemical features distinguishing neuroendocrine carcinomas of the larynx from paraganglioma, metastatic medullary carcinoma, malignant melanoma, and basaloid squamous cell carcinoma are presented. The second portion of this review outlines the clinicopathologic features of two head and neck neoplasms exhibiting varying degrees of neuroendocrine differentiation. Olfactory neuroblastomas have well-developed neuroendocrine differentiation, almost invariably arise from the olfactory mucosa, typically exhibit low-grade cytologic features, and may have protracted clinical course with an approximately 50% overall 5-yr survival. In contrast, sinonasal undifferentiated carcinoma is a microscopically high-grade neoplasm with minimal, abortive neuroendocrine features, a highly aggressive clinical course, and virtually 100% mortality. They can arise throughout the sinonasal region. Presented at the Endocrine Pathology Society—USCAP Meeting, Washington, DC, March 23, 1996.     

Growth factors in melanoma

Human melanoma cells in culture are the source of a wide variety of polypeptide growth factors. Melanoma-derived basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF)-A and PDGF-B chains, transforming growth factor (TGF)- and TGF-, interleukin (IL)-1 and IL-1, and melanoma growth stimulatory activity (MGSA) have similar biochemical and functional properties when compared to their counterparts produced by untransformed cells. In contrast to melanoma cells, normal melanocytes, even under optimal growth conditions, express only TGF-1 and MGSA at detectable levels suggesting that production of the other growth factors is a tumor-associated phenomenon. Recent evidence suggests that at least two of the growth factors, bFGF and MGSA, contribute to autocrine growth stimulation of melanoma cells. Whether PDGF, TGF-, IL-1, and TGF- act in an autocrine mode is unclear at present. However, these four growth factors are among those secreted by melanoma cells and, therefore, can be expected to interact with normal cells of the tumor stroma in vivo. Such paracrine effects include not only growth modulation in the context of angiogenesis and stroma formation, but also tissue degradation by proteolytic enzymes, the modification of extracellular matrix composition, and expression of adhesion receptors.     

Structure,function and biological properties of integrin αvβ3on human melanoma cells

Summary Human melanoma represents one of the most metastatic cancers in man. The capacity of melanoma cells to invade a variety of tissues and extracellular matrices is, in part, due to their repertoire of adhesion receptors. To this end, human melanoma cells express multiple integrin cell adhesion receptors among these is the vitronectin receptor, _v3. This adhesion receptor enables melanoma cells to attach to a wide variety of extracellular matrix components containing the sequence Arg-Gly-Asp. This review will focus on the biosynthetic, biochemical and biological properties of this receptor expressed on the surface of human melanoma cells.     

In vivo evaluation of cell mediated immunity against human melanoma

Evaluation of cell mediated immunity against human melanoma target cells was performed in an in vivo model using human tumor xenografts growing in Balb/c athymic mice. Intraperitoneal inoculation of 1 × 10⁷ human melanoma cells produced peritoneal carcinomatosis which lead to death of the animals at 23.8 ± 2.6 days (N = 12). Peripheral blood lymphocytes (PBL) from normal donors were administered to tumor bearing mice, and survival times of 22.0±2.3 days were observed (N = 8). Peripheral blood lymphocytes from four of five normal donors which had been presensitized on monolayers of melanoma tissue culture cells in vitro failed to prolong host survival times. In contrast, PBL obtained from 15 of 20 melanoma patients were found to prolong survival of the tumor bearing nude mice. Of these 15 patients, 8 were undergoing specific active immunotherapy, while 7 had not been sensitized except by the disease process. The ability of PBL obtained from patients to prolong survival of tumor bearing animals did not appear to correlate with either the stage of the disease or the patient's clinical course. The possible mechanisms for the prolonged survival and usefulness of this model are discussed.