NRF3 Decreases during Melanoma Carcinogenesis and Is an Independent Prognostic Marker in Melanoma

The prognostic significance of the major redox regulator, nuclear factor erythroid-2-related factor 2 (NRF2), is recognized in many cancers, but the role of NRF3 is not studied. Analysis from the Gene Expression Omnibus datasets showed that NRF3 mRNA levels increased from benign to dysplastic naevi (p = 0.04). We characterized the immunohistochemical expression of NRF3 in 81 naevi, 67 primary skin melanomas, and 51 lymph node metastases. The immunohistochemical expression of cytoplasmic NRF3 decreased from benign to dysplastic naevi (p < 0.001) and further to primary melanomas (p < 0.001). High cytoplasmic NRF3 protein expression in pigment cells of the primary melanomas associated with worse melanoma-specific survival in multivariate analysis, specifically in the subgroup of patients with the lymph node metastases at the time of diagnosis (hazard ratio 3.179; 95% confidence interval 1.065-9.493; p = 0.038). Intriguingly, we did not observe associations between NRF3 and the traditional prognostic factors such as Breslow thickness, ulceration, or stage. Together, this data represents the primary description about the role of NRF3 in pigment tumours that is worthy of further explorations.Peer reviewe

The Transcription Factor Twist1 Has a Significant Role in Mycosis Fungoides (MF) Cell Biology: An RNA Sequencing Study of 40 MF Cases

The purpose of this RNA sequencing study was to investigate the biological mechanism underlying how the transcription factors (TFs) Twist1 and Zeb1 influence the prognosis of mycosis fungoides (MF). We used laser-captured microdissection to dissect malignant T-cells obtained from 40 skin biopsies from 40 MF patients with stage I–IV disease. Immunohistochemistry (IHC) was used to determinate the protein expression levels of Twist1 and Zeb1. Based on RNA sequencing, principal component analysis (PCA), differential expression (DE) analysis, ingenuity pathway analysis (IPA), and hub gene analysis were performed between the high and low Twist1 IHC expression cases. The DNA from 28 samples was used to analyze the TWIST1 promoter methylation level. In the PCA, Twist1 IHC expression seemed to classify cases into different groups. The DE analysis yielded 321 significant genes. In the IPA, 228 significant upstream regulators and 177 significant master regulators/causal networks were identified. In the hub gene analysis, 28 hub genes were found. The methylation level of TWIST1 promoter regions did not correlate with Twist1 protein expression. Zeb1 protein expression did not show any major correlation with global RNA expression in the PCA. Many of the observed genes and pathways associated with high Twist1 expression are known to be involved in immunoregulation, lymphocyte differentiation, and aggressive tumor biology. In conclusion, Twist1 might be an important regulator in the disease progression of MF

Upregulated integrin α11 in the stroma of cutaneous squamous cell carcinoma promotes skin carcinogenesis

Integrin α11β1 is a collagen-binding integrin that is needed to induce and maintain the myofibroblast phenotype in fibrotic tissues and during wound healing. The expression of the α11 is upregulated in cancer-associated fibroblasts (CAFs) in various human neoplasms. We investigated α11 expression in human cutaneous squamous cell carcinoma (cSCC) and in benign and premalignant human skin lesions and monitored its effects on cSCC development by subjecting α11-knockout (Itga11−/−) mice to the DMBA/TPA skin carcinogenesis protocol. α11-deficient mice showed significantly decreased tumor cell proliferation, leading to delayed tumor development and reduced tumor burden. Integrin α11 expression was significantly upregulated in the desmoplastic tumor stroma of human and mouse cSCCs, and the highest α11 expression was detected in high-grade tumors. Our results point to a reduced ability of α11-deficient stromal cells to differentiate into matrix-producing and tumor-promoting CAFs and suggest that this is one causative mechanism underlying the observed decreased tumor growth. An unexpected finding in our study was that, despite reduced CAF activation, the α11-deficient skin tumors were characterized by the presence of thick and regularly aligned collagen bundles. This finding was attributed to a higher expression of TGFβ1 and collagen crosslinking lysyl oxidases in the Itga11-/- tumor stroma. In summary, our data suggest that α11β1 operates in a complex interactive tumor environment to regulate ECM synthesis and collagen organization and thus foster cSCC growth. Further studies with advanced experimental models are still needed to define the exact roles and molecular mechanisms of stromal α11β1 in skin tumorigenesis.publishedVersio

Reactive oxygen species-mediated mechanisms of action of targeted cancer therapy

Abstract Targeted cancer therapies, involving tyrosine kinase inhibitors and monoclonal antibodies, for example, have recently led to substantial prolongation of survival in many metastatic cancers. Compared with traditional chemotherapy and radiotherapy, where reactive oxygen species (ROS) have been directly linked to the mediation of cytotoxic effects and adverse events, the field of oxidative stress regulation is still emerging in targeted cancer therapies. Here, we provide a comprehensive review regarding the current evidence of ROS-mediated effects of antibodies and tyrosine kinase inhibitors, use of which has been indicated in the treatment of solid malignancies and lymphomas. It can be concluded that there is rapidly emerging evidence of ROS-mediated effects of some of these compounds, which is also relevant in the context of drug resistance and how to overcome it

Optimization of measurement mode and sample processing for FTIR microspectroscopy in skin cancer research

Abstract The use of Fourier Transform Infrared (FTIR) microspectroscopy to study cancerous cells and tissues has gained popularity due to its ability to provide spatially resolved information at the molecular level. Transmission and transflection are the commonly used measurement modes for FTIR microspectroscopy, and the tissue samples measured in these modes are often paraffinized or deparaffinized. Previous studies have shown that variability in the spectra acquired using different measurement modes and sample processing methods affect the result of the analysis. However, there is no protocol that standardizes the mode of measurement and sample processing method to achieve the best classification result. This study compares the spectra of primary (IPC-298) and metastatic (SK-MEL-30) melanoma cell lines acquired in both transmission and transflection modes using paraffinized and deparaffinized samples to determine the optimal combination for accurate classification. Significant differences were observed in the spectra of the same cell line measured in different modes and with or without deparaffinization. The PLS-DA model built for the classification of two cell lines showed high accuracy in each case, suggesting that both modes and sample processing alternatives are suitable for differentiating cultured cell samples using supervised multivariate analysis. The biochemical information contained in the cells capable of discriminating two melanoma cell lines is present regardless of mode or sample type used. However, the paraffinized samples measured in transflection mode provided the best classification

Discrimination of melanoma cell lines with Fourier Transform Infrared (FTIR) spectroscopy

Abstract Among skin cancers, melanoma is the lethal form and the leading cause of death in humans. Melanoma begins in melanocytes and is curable at early stages. Thus, early detection and evaluation of its metastatic potential are crucial for effective clinical intervention. Fourier transform infrared (FTIR) spectroscopy has gained considerable attention due to its versatility in detecting biochemical and biological features present in the samples. Changes in these features are used to differentiate between samples at different stages of the disease. Previously, FTIR spectroscopy has been mostly used to distinguish between healthy and diseased conditions. With this study, we aim to discriminate between different melanoma cell lines based on their FTIR spectra. Formalin-fixed paraffin embedded samples from three melanoma cell lines (IPC-298, SK-MEL-30 and COLO-800) were used. Statistically significant differences were observed in the prominent spectral bands of three cell lines along with shifts in peak positions. A partial least square discriminant analysis (PLS-DA) model built for the classification of three cell lines showed an overall accuracy of 92.6% with a sensitivity of 85%, 95.75%, 96.54%, and specificity of 97.80%, 92.14%, 98.64% for the differentiation of IPC-298, SK-MEL-30, and COLO-800, respectively. The results suggest that FTIR spectroscopy can differentiate between different melanoma cell lines and thus potentially characterize the metastatic potential of melanoma

The use of Fourier Transform Infrared (FTIR) spectroscopy in skin cancer research:a systematic review

Abstract Skin cancers are one of the most frequently occurring diseases in humans that pose severe health issues. Fourier Transform Infrared (FTIR) spectroscopy in cancer research has gained considerable attention because of its ability to provide biochemical information in addition to being compatible with traditional histopathology. With this review, we aim to identify all skin cancer studies which have been conducted using FTIR spectroscopy and depict different methodologies that have been used to analyze FTIR spectroscopic data of skin cancers. We conducted the systematic review following PRISMA guidelines for which three databases, Scopus, PubMed and Web of Science, were searched from commencement to 16 January 2019. All the studies which used FTIR spectroscopy for skin cancer research were included in the review. A total of 35 studies were found eligible to be included in the review. Of these, 21 studies were based on melanoma, 6 studies on BCC, 2 studies on SCC, and 2 on lymphocytes. The remaining 4 studies aimed to differentiate between various skin cancer types. The potential of FTIR spectroscopy for many relevant aspects of skin cancer research has already been demonstrated, but more work is needed to establish FTIR spectroscopy as a routine method in the field

NRF3 decreases during melanoma carcinogenesis and Is an independent prognostic marker in melanoma

Abstract The prognostic significance of the major redox regulator, nuclear factor erythroid-2-related factor 2 (NRF2), is recognized in many cancers, but the role of NRF3 is not studied. Analysis from the Gene Expression Omnibus datasets showed that NRF3 mRNA levels increased from benign to dysplastic naevi (p = 0.04). We characterized the immunohistochemical expression of NRF3 in 81 naevi, 67 primary skin melanomas, and 51 lymph node metastases. The immunohistochemical expression of cytoplasmic NRF3 decreased from benign to dysplastic naevi (p &lt; 0.001) and further to primary melanomas (p &lt; 0.001). High cytoplasmic NRF3 protein expression in pigment cells of the primary melanomas associated with worse melanoma-specific survival in multivariate analysis, specifically in the subgroup of patients with the lymph node metastases at the time of diagnosis (hazard ratio 3.179; 95% confidence interval 1.065-9.493; p = 0.038). Intriguingly, we did not observe associations between NRF3 and the traditional prognostic factors such as Breslow thickness, ulceration, or stage. Together, this data represents the primary description about the role of NRF3 in pigment tumours that is worthy of further explorations

NRF1 and NRF2 mRNA and protein expression decrease early during melanoma carcinogenesis:an insight into survival and microRNAs

Abstract The prognostic significance of the major redox regulator nuclear factor erythroid-2-related factor (NRF2) is recognized in many cancers, but the role of NRF1 is not generally well understood in cancer. Our aim was to investigate these redox transcription factors in conjunction with redox-related microRNAs in naevi and melanoma. We characterized the immunohistochemical expression of NRF1 and NRF2 in 99 naevi, 88 primary skin melanomas, and 67 lymph node metastases. In addition, NRF1 andNRF2 mRNA and miR-23B, miR-93, miR-144, miR-212, miR-340, miR-383, and miR-510 levels were analysed with real-time qPCR from 54 paraffin-embedded naevi and melanoma samples. The immunohistochemical expression of nuclear NRF1decreased from benign to dysplastic naevi (p &lt; 0:001) and to primary melanoma (p&lt;0:001) and from primary melanoma to metastatic lesions (p=0:012). Also, NRF1 mRNA levels decreased from benign naevi to dysplastic naevi (p=0:034). Similarly, immunopositivity of NRF2 decreased from benign to dysplastic naevi (p=0:02) and to primary lesions (p=0:018). NRF2mRNA decreased from benign to dysplastic naevi and primary melanomas (p=0:012). Analysis from the Gene Expression Omnibus datasets supported the mRNA findings. High nuclear immunohistochemical NRF1 expression in pigment cells associated with a worse survival (p=0:048) in patients with N0 disease at the time of diagnosis, and high nuclear NRF2 expression in pigment cells associated with a worse survival (p=0:033) in patients with M0 disease at the time of diagnosis. In multivariate analysis, neither of these variables exceeded the prognostic power of Breslow. The levels of miR-144 and miR-212 associated positively with ulceration (p=0:012 and p=0:027, respectively) while miR-510 levels associated positively with lymph node metastases at the time of diagnosis (p=0:004). Furthermore, the miRNAs correlated negatively with the immunohistochemical expression of NRF1 and NRF2 but positively with their respective mRNA. Together, this data sheds new light about NFE2L family factors in pigment tumors and suggests that these factors are worth for further explorations