Chromosomal aberrations in squamous cell carcinoma and solar keratoses revealed by comparative genomic hybridization

OBJECTIVE: To identify chromosomal copy numbers of frequent genetic aberrations within squamous cell carcinomas (SCCs) and solar keratoses (SKs), and provide further evidence to support or challenge current dogma concerning the relationship between these lesions. DESIGN: Retrospective analysis of genetic aberrations in DNA from SK and SCC biopsy specimens by comparative genomic hybridization. SETTING: University-based research laboratory in Queensland, Australia. PATIENTS: Twenty-two biopsy specimens from patients with diagnosed SKs (n = 7), cutaneous SCCs (n = 10), or adjoining lesions (n = 5). MAIN OUTCOME MEASURES:Identification of frequent genetic aberrations both specific to SK and SCC and shared by these lesions to investigate their clonal relationship. RESULTS: Shared genomic imbalances were identified in SK and SCC. Frequent gains were located at chromosome arms 3q, 17q, 4p, 14q, Xq, 5p, 9q, 8q, 17p, and 20q, whereas shared regional losses were observed at 9p, 3p, 13q, 17p, 11p, 8q, and 18p. Significant loss of 18q was observed only in SCC lesions. CONCLUSIONS: Our results demonstrate that numerous chromosomal aberrations are shared by the 2 lesions, suggesting a clonal relationship between SK and SCC. Additionally, the genomic loss of 18q may be a significant event in SK progression to SCC. Finally, the type and frequency of aberrations suggests a common mode of tumorigenesis in SCC-derived tumors.     

Array comparative genomic hybridization of keratoacanthomas and squamous cell carcinomas: different patterns of genetic aberrations suggest two distinct entities

Keratoacanthoma (KA) is a benign keratinocytic neoplasm that spontaneously regresses after 3-6 months and shares features with squamous cell carcinomas (SCCs). Furthermore, there are reports of KAs that have metastasized, invoking the question of whether KA is a variant of SCC (Hodak et al., 1993). To date, no reported criteria are sensitive enough to discriminate reliably between KA and SCC, and consequently there is a clinical need for discriminating markers. Our previous study analyzed 132 KAs and 29 SCCs and revealed significantly different regions of genomic aberrations using chromosomal comparative genomic hybridization (CGH). In the present study, we applied array CGH to investigate 98 KAs and 22 SCCs from the above samples. The result shows that all KAs and SCCs have some degree of genetic aberrations. The distribution of numbers of aberrant clones per sample differed significantly between KAs and SCCs (P<0.02), which also demonstrated recurrent aberrations that differed significantly (P<0.001), as illustrated by unsupervised cluster analysis. Classifiers for clinicopathological parameters of KAs were established based on t-test statistics and permutation tests. Tumor size, fibrosis, and inflammation, which are related to the developmental stages of KAs, showed significant (t-test, permutation test) associations with aberrations of selected genomic regions. This suggests chromosomal instability during the whole life cycle of KAs.     

Detection of numerical chromosomal aberrations in malignant melanomas using fluorescence in situ hybridization

To evaluate the numerical chromosomal aberration i malignant melanoma, we have applied fluorescence in situ hybridization (FISH) with repetitive DNA probes specific for chromosomes 1, 6, 7, 9, 10, and 17 on 24 fresh malignant melanomas (primary: 14, metastatic: 8).
We defined a tumor that had copies with more than 3 spots as chromosomal gain. Chromosomal copy number gain was found in 40.9% of the cases for chromosome 7, 27.2% for chromosome 6, 27.2% for chromosome 17, 22.7% for chromosome 9 and 10, and 4.5% for chromosome I. Monosomy was found in 54.5% of the cases for chromosome 10, 36.5% for chromosome 9, 27.2% for chromosome 6, 22.7% for chromosome17, and 18.1% for chromosome 1 and 7 The most frequent numerical alterations were seen in chromosomes 6, 7, 9 and 10.
Gain of chromosome 6 and 7 and/or losses of chromosome 9 and 10 may play an important role in the tumorigenesis and development of malignant melanomas.     

Analysis of cytogenetic abnormalities in squamous cell carcinoma by array comparative genomic hybridization

IntroductionFew conventional cytogenetic studies of squamous cell carcinoma (SCC) have been performed to date. The introduction of cytogenetic techniques such as comparative genomic hybridization (CGH) has resolved some of the problems associated with conventional cytogenetics. The aim of this study was to analyze the presence of genetic abnormalities in a series of patients with SCC using the technique of array CGH.Material and methodsThe study included 8 patients (7 men and 1 woman; mean age, 75 years) diagnosed with primary SCC. DNA was extracted from frozen tissue and analyzed by array CGH.ResultsAll cases had genetic alterations, with gains more frequent than losses. The chromosomal regions with gains, in descending order of frequency, were as follows: 5p15.2, 9q31.3-q33.2, 13q, 18q22, 1p21-p22, 1q24-q25, 3p13, 4q33-q34 (HMGB2, SAP30), 20p12.2 (JAG1), 21q21.1, and Xq21.33. The region 9p13.1-p13.3 was the only one to display recurrent loss. No correlation was observed between the presence of gains or losses and the clinical and pathological characteristics of the tumors.ConclusionsThis is the first study to use the technique of array CGH to analyze genetic alterations in SCC. The finding of certain previously described aberrations (gain of 5p) suggests the existence of recurrent abnormalities. Likewise, the observation of alterations in small regions of chromosome 1 highlights the sensitivity of the technique to detect small changes. Application of the technique to a larger series of cases will provide greater insight into the genetic abnormalities implicated in the process of tumorigenesis in SCC.     

Distinguishing melanocytic nevi from melanoma by DNA copy number changes: comparative genomic hybridization as a research and diagnostic tool

Cancer typically results in loosened control over genomic integrity, resulting in alterations of the genome of cancer cells. Comparative genomic hybridization (CGH) is a method that can be used on DNA extracted from routinely fixed tissue to assess the entire genome for the presence of changes in DNA copy number. CGH analysis has revealed that melanoma differs from melanocytic nevi by the presence of frequent chromosomal aberrations. In contrast, melanocytic nevi typically show no chromosomal aberrations, or have a restricted set of alterations with basically no overlap to melanoma. These marked differences between aberration patterns in melanomas and melanocytic nevi can be exploited diagnostically to classify melanocytic tumors that are ambiguous based on histopathologic assessment. In addition to potential diagnostic applications, detailed analyses of recurrent aberrations can lead to the identification of genes relevant in melanocytic neoplasia.     

Molecular cytogenetic analysis of basal cell carcinoma DNA using comparative genomic hybridization

In an attempt to define genomic copy number changes associated with the development of basal cell carcinoma, we investigated 15 sporadic tumors by comparative genomic hybridization. With the incorporation of tissue microdissection and degenerate oligonucleotide primed-polymerase chain reaction we were able to isolate, and then universally amplify, DNA from the tumor type. This combined approach allows the investigation of chromosomal imbalances within a histologically distinct region of tissue. Using comparative genomic hybridization we have observed novel and recurrent chromosomal gains at 6p (47%), 6q (20%), 9p (20%), 7 (13%), and X (13%). In addition comparative genomic hybridization revealed regional loss on 9q in 33% of tested tumors encompassing 9q22.3 to which the putative tumor suppressor gene, Patched, has been mapped. The deletion of Patched has been indicated in the development of hereditary and sporadic basal cell carcinomas. The identification of these recurrent genetic aberrations suggests that basal cell carcinomas may not be as genetically stable as previously thought. Further investigation of these regions may lead to the identification of other genes responsible for basal cell carcinoma formation.     

Proximal-type epithelioid sarcoma: case report and result of comparative genomic hybridization

BACKGROUND: Epithelioid sarcoma is a rare mesenchymal neoplasm. Recently, a more aggressive, so-called "proximal type" epithelioid sarcoma has been described. CLINICAL CASE: A 40-year-old-woman presented with 5 x 4 cm, erythematous, indurated, non-movable, painful mass on the pubic area. Histopathology demonstrated diffuse tumor-cell infiltration into the subcutaneous and fascia, which was consisted of prominent epithelioid cells and scattered rhabdoid cells. A multinodular growth pattern or granulomatous appearance with central necrosis was not observed. The tumor cells showed positive reactions for vimentin, cytokeratin (AE1/AE3), and CD34. Despite the surgery, left inguinal mass with lymphadenopathy occurred one month later. We also carried out comparative genomic hybridization (CGH) with tumor cells. CGH revealed chromosomal gain of 5q32-qter, 12q24-qter, and 22q. CONCLUSION: We report a case of proximal-type of epithelioid sarcoma, which showed the chromosomal gains of 5q32-qter, 12q24-qter, and 22q by CGH.     

A case of adult T-cell leukaemia/lymphoma characterized by multiplex-fluorescence in situ hybridization, comparative genomic hybridization, fluorescence in situ hybridization and cytogenetics

Adult T-cell leukaemia/lymphoma (ATLL) is a neoplasm of mature helper (CD4) T lymphocytes. Little is known, however, about the chromosome aberrations associated with the pathogenesis of this malignancy. Using molecular cytogenetic techniques we, therefore, investigated a 44-year-old man who had a 7-year history of ATLL with cutaneous involvement mimicking primary cutaneous T-cell lymphoma. Conventional cytogenetics revealed gross chromosomal changes with chromosome numbers ranging from 71 to 82. There were structural abnormalities of chromosomes 7 and 9, partial deletions of chromosomes 1, 3, 5 and 6, and loss of chromosomes 2, 4, 9, 11–14, 21 and 22. Multiplex-fluorescence in situ hybridization (M-FISH) identified two derivative chromosomes, der(6)t(6;7)(q16;q21) and der(7)t(6;7)(q16;q21)ins(6;12)(q2?;?), and a deletion of chromosome 1p. Conventional FISH confirmed the M-FISH findings. Comparative genomic hybridization of the blood revealed gains of DNA copy number at 1q12–25, 6p24–25, 9p23, 16p13–q13, 17q11–21, 19p13 and 20q13 and loss at 11p15 while lymph nodes showed gains at 3p22–24, 3q27–29, 7q36 and 15q26 and losses at 2p24–25, 2q37, 10p14–15, 11p15, 13q33–34 and 16p13.3. No DNA copy number changes were seen in a skin lesion. These results show the extent of genetic abnormalities within this malignancy.     

Comparative genomic hybridization (CGH) discloses chromosomal and subchromosomal copy number changes in Merkel cell carcinomas

We analyzed three Merkel cell carcinomas (MCC), applying comparative genomic hybridization (CGH) with DNA from paraffin-embedded and cultured tumor material as the probes. By this method, numerous changes in chromosome copy numbers were observed in each tumor investigated. Recurrent gains of chromosomes 1, 6, 18q and 20 were detected in two tumors. A third tumor showed complex chromosomal copy number changes, including gain of chromosome 8 and 9. These gains, as well as gain of chromosome 1 in the first two tumors, were confirmed by fluorescence in situ hybridization to paraffin tissue sections. Our results support the view that important genes for MCC development may be located on chromosomes 1, 6, 18q and 20.     

Microarray comparative genomic hybridization detection of copy number changes in desmoplastic melanoma and malignant peripheral nerve sheath tumor

Desmoplastic melanoma (DM) and malignant peripheral nerve sheath tumor (MPNST) can appear morphologically and immunophenotypically similar. We attempted to determine whether microarray comparative genomic hybridization could detect copy number differences between them to aid in the diagnosis. S-100 immunohistochemistry was performed on 5 cases of DM and 9 cases of MPNST using formalin-fixed paraffin-embedded specimens. Genomic DNA was extracted from microdissected cells. Whole genome amplification was performed on 5 of 5 DMs and 6 of 9 MPNST cases. A multiplex polymerase chain reaction assay was used to determine the quality of the DNA samples, which were run on the Spectral Chip 2600 bacterial artificial chromosome array platform. DM showed gains involving chromosomes 1p, 2p, 9q, 13q, 14q, and 20q and losses involving chromosomes 5p, 11p, 12q, 15q, and 18q. Several cancer-associated genes were involved, including gain of BCL2L1, ARTN, AMPK, NRAS, and CCNA1 and loss of IGF2, CDKN1C, PAX6, WT1, TRAF6, MAPK8IP1, and IMP3. MPNST had gains involving chromosomes 1p, 2q, and 19p and loss of chromosome 21q. Gains of MUM1, APC2, MAP2K2, JMJD2B, SP110, PTMA, GPI, and CDKN2D were detected. DM and MPNST have chromosomal alterations detected by array comparative genomic hybridization that might be useful in distinguishing these 2 tumors, although further studies with a larger sample size will be needed to test this.     

Basal cell carcinoma of the skin: whole genome screening by comparative genome hybridization revisited

Basal cell carcinoma (BCC) of the skin is considered to be the most common malignancy in people of European ancestry. It is often not clinically aggressive and has been regarded as genetically stable. However, histopathologic subtypes of BCC differ in their ability to invade surrounding tissues and recur. The aim of this work was to present a comprehensive study of chromosomal imbalances of cutaneous BCC and to correlate the findings with their histopathologic and clinical features. In all, 101 tumor samples were classified according to the current microscopic classification of BCC and then analyzed by comparative genomic hybridization (CGH). Over 60% of BCC were found to carry chromosomal imbalances – partial or whole chromosome gains and losses. Different subtypes of BCC presented common chromosomal alterations. No single chromosomal imbalance was found to be characteristic of a particular subtype of BCC, although the frequency of some chromosomal changes differed from one group to the other. The significance of chromosome 2 gains as a phenomenon that does not coexist with the losses in 9q is discussed.     

Primary dapsone resistance as assessed by uptake of labelled thymidine by the macrophage resident Mycobacterium leprae

12 untreated lepromatous leprosy patients were screened for primary dapsone resistance by the uptake of labelled thymidine by macrophage resident M. leprae. There were found to harbour primary dapsone resistant strains of M. leprae and another three partially resistant strains to the drug. This rapid, simple and reliable method should be used routinely to screen leprosy patients, for drug resistance.     

Thickness and echogenicity of the skin in children as assessed by 20-MHz ultrasound

BACKGROUND: Skin anatomy and physiology undergo modifications throughout the whole lifespan. In children the skin appears with structural characteristics, as evaluated by histology, which differ from those of adults, especially in the first years of life. OBJECTIVE: The aim of our study was to evaluate skin thickness and echogenicity at different sites in children by 20-MHz ultrasound. METHODS: Eight skin sites of 42 children and 30 young adults were studied with a 20-MHz B scanner. Skin thickness and mean echogenicity were evaluated. The echographic images were processed and segmented by a dedicated program: the 0-30 amplitude interval, which marks the hypoechogenic parts of the tissue, and the 201-255 range, highlighting the hyperreflecting areas, have been selected. RESULTS AND CONCLUSION: Whereas skin thickness shows a gradual increase from birth to adulthood, maturation of the skin leads to variations in the intensity of its echogenicity, depending on the different skin areas. Whereas on the face and the trunk it appears lower in adults with respect to children, a gradual increase can be observed on the limbs with growing age. The distribution of skin reflectivity also greatly varies in different phases of life.     

Oxygen levels in normal and previously irradiated human skin as assessed by EF5 binding

The oxygen status of skin is a controversial topic. Skin is radiosensitive, suggesting it is well-oxygenated. However, it can be further sensitized with nitroimidazole drugs, implying that it is partially hypoxic. Skin oxygen levels are difficult to measure with either electrodes or the hypoxia-monitoring agent (3)H-misonidazole. For the latter, binding has previously been reported to be high in murine skin, but this could be attributed to either non-oxygen-dependent variations in nitroreductase activity, drug metabolism, and/or actual oxygen gradients. We obtained tumor and skin from patients given EF5, a 2-nitroimidazole tissue hypoxia monitor. We performed immunohistochemical studies using highly specific monoclonal antibodies for the hypoxia-dependent production of EF5 tissue adducts. Some tissue sections were counterstained using either Ki67 for proliferation or CD31 for vessels. We found that the human dermis is well-oxygenated, the epidermis is modestly hypoxic and portions of some sebaceous glands and hair follicles are moderately to severely hypoxic. Normal and irradiated skin had similar oxygenation patterns. Control studies demonstrated that these observations are not due to tissue variations in nitroreductase activity. The importance of the highly heterogeneous distribution of oxygen in skin requires further study, but recent investigations suggest that skin hypoxia may have important clinical ramifications including mediating cellular transformation.     

Life quality of Chinese patients with chronic urticaria as assessed by the dermatology life quality index

Background No report focused on the life quality of Chinese patients with chronic urticaria (CU). It was also due to lack of data to assess the usage of the Dermatology Life Quality Index (DLQI) in Chinese patients. Objectives We explored the quality of life (QoL) of Chinese patients with CU and assessed the factor structure and reliability of the Chinese version of the DLQI. Methods The DLQI scores were recorded to evaluate the life quality of out‐patients with CU. The reliability and the factor structure of the DLQI were assessed using reliability analysis and factor analysis. Results A total of 131 patients completed the DLQI. The mean score was 9.93 that was significantly associated with the patient’s age (P = 0.01). Chronic urticaria had the greatest effect on the ‘Syndrome and Feeling’ (50.64%), followed by ‘Work/School’ (41.98%). A reliability analysis showed that the value of the Cronbach’sα reached 0.85 if item 1 in the DLQI was excluded. Both exploratory and confirmatory factor analysis indicated that there were two latent common factors underlying the items in the questionnaire. Conclusions Chronic urticaria has a moderate impact on the life quality. More attention should be paid to the interference from item 1 in assessing the QoL by the DLQI, although it remains a concise tool with high reliability.     

The regional distribution of neuropeptides in human skin as assessed by radioimmunoassay and high-performance liquid chromatography

In this study radioimmunoassay was used to determine neuropeptide levels in extracts from 17 differing anatomical regions of human skin. Marked regional variations of neuropeptide content for human skin were found and these variations are likely to reflect true physiological functions for the neuropeptides studied. In general the tachykinins, substance P (SP), neurokinin A (NKLA) and calcitonin gene-related peptide (CGRP) were found in highest concentrations in regions of skin with the greatest tactile sensation. By contrast, highest concentrations of vasoactive intestinal peptide (VIP) and peptide histidine methionine (PHM) were found in axillary skin, where they probably play a part in axillary eccrine sweat production. Neurotensin was not found in any of the skin areas sampled, suggesting that it is relatively unimportant in human physiological skin control. Reverse-phase high-performance liquid chromatography (rpHPLC) was used to verify the results of radioimmunoassay. Both SP and NKA occurred in several regions in both their reduced and oxidized forms, as well as displaying molecular heterogeneity. CGRP occurred as one molecular species, this being α-CGRP, suggesting that this is the predominant molecular form in human skin. Likewise, both VIP and PHM displayed molecular homogeneity in the regions investigated by rpHPLC.     

Lipophilic and hydrophilic moisturizers show different actions on human skin as revealed by cryo scanning electron microscopy

To study the mode of action of moisturizers on human skin, hydrophilic moisturizers in water and neat lipophilic moisturizers were applied on excised skin for 24 h at 32 degrees C. Samples of the treated skin were subsequently visualized in a cryoscanning electron microscope. The stratum corneum (SC) appeared as a region of swollen corneocytes (the swollen region) sandwiched between two layers of relatively dry corneocytes (the upper and lower non-swelling regions respectively). Lipophilic moisturizers increased the water content of the SC, whereas hydrophilic moisturizers can also reduce the water content of the SC. When focusing on the effect of the moisturizers on the three different regions, it was observed that cells in the swelling region are most sensitive to the application of the moisturizers and that the change in SC thickness is most influenced by the change in the thickness of the swelling region. Summarizing, SC cells are not equally sensitive to moisturizer application: centrally located corneocytes are more sensitive than corneocytes in the upper and the lowest regions of the SC.     

Transient improvement of urticaria induces poor adherence as assessed by Morisky Medication Adherence Scale‐8

Poor adherence to medication is a major public health challenge. Here, we aimed to determine the adherence to oral and topical medications and to analyze underlying associated factors using the translated Japanese version of Morisky Medication Adherence Scale‐8 regarding urticaria treatment. Web‐based questionnaires were performed for 3096 registered dermatological patients, along with a subanalysis of 751 registered urticaria patients in this study. The adherence to oral medication was significantly associated with the frequency of hospital visits. Variables that affected the adherence to topical medication included age and experience of drug effectiveness. The rate of responses that “It felt like the symptoms had improved” varied significantly among the dermatological diseases treated with oral medications. Dermatologists should be aware that adherence to the treatment of urticaria is quite low. Regular visits and active education for patients with urticaria are mandatory in order to achieve a good therapeutic outcome by increasing the adherence.