Detection of JunB and C-FLIP Psoriasis Genes Using the Reverse Transcriptase in-situ PCR Technique
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Abstract
Objective Psoriasis is a chronic skin disease and considered a multifactorial condition as it is caused by several risk factors; including genetic and environmental interactions. This study aimed to examine localization the expression of JunB and c-FLIP psoriasis genes at the mRNA cellular level using reverse transcriptase in-situ PCR (in-situ RT-PCR).
Methods The in-situ RT-PCR technique was used to amplify the targeted mRNA from the formalin-fixed and paraffin-embedded tissues of three psoriasis patients. The localization of JunB and c-FLIP mRNA expression was achieved using in-situ RT-PCR along with the immunodetection technique. An immunohistochemistry profiler was used to count the pixels including the percentage contributions.
Results Hematoxylin and eosin staining revealed the presence of parakeratosis, acanthosis, spongiosis and inflammatory cells. The staining analysis outcomes from in-situ RT-PCR and immunohistochemistry detection showed that the c-FLIP genes and JunB genes were significantly expressed in the surrounding nucleus area at the cellular level of the psoriasis tissue. JunB and c-FLIP mRNA scored positively in the the immunohistochemical analysis.
Conclusions RT in-situ PCR can be a tool for identifying mRNA cellular gene expression. JunB and c-FLIP gene expression are significantly present in psoriasis.
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References
Boehncke W, Schon M. Psoriasis. Lancet. 2015;386: 983-4.
Capon F. The Genetic Basis of Psoriasis. Int J Mol Sci. 2017;18:2526. PubMed PMID: 29186830
Yan D, Gudjonsson J, Le S, Maverakis E, Plazyo O, Ritchlin C, et al. New frontiers in psoriatic disease research, part I: genetics, environmental triggers, immunology, pathophysiology, and precision medicine. J Invest Dermatol. 2021;141:2112-22.
Tsoi L, Stuart P, Tian C, Gudjonsson J, Das S, Zawistowski M, et al. Large scale meta-analysis characterizes genetic architecture for common psoriasis associated variants. Nat Commun. 2017;8:15382. PubMed PMID: 28537254
Nair R, Duffin K, Helms C, Ding J, Stuart P, Goldgar D, et al. Genome-wide scan reveals association of psoriasis with IL-23 and NF-B pathways. Nat Genet. 2006;38:484-91.
Balendran N, Clough R, Arguello J, Barber R, Veal C, Jones A, et al. Characterization of the major susceptibility region for psoriasis at chromosome 6p21.3. J Invest Dermatol. 1999;113:322-8.
Bowcock A, Krueger J. Getting under the skin: the immunogenetics of psoriasis. Nat Rev Immunol. 2005;5: 699-711.
Griffiths C, Barker J. Pathogenesis and clinical features of psoriasis. Lancet. 2007;370:263-71.
Mehic D, Bakiri L, Ghannadan M, Wagner E, Tschachler E. Fos and jun proteins are specifically expressed during differentiation of human keratinocytes. J Invest Dermatol. 2005;124:212-20.
Piechaczyk M, Farràs R. Regulation and function of JunB in cell proliferation. Biochem Soc Trans. 2008; 36:864-7.
Zenz R, Eferl R, Kenner L, Florin L, Hummerich L, Mehic D, et al. Psoriasis-like skin disease and arthritis caused by inducible epidermal deletion of Jun proteins. Nature. 2005;437:369-75.
Yang J, Li Y, Liu YQ, Long JW, Tian F, Dong J, et al. Expression of antiapoptotic protein c-FLIP is upregulated in psoriasis epidermis. Eur J Dermatol. 2009;19:29-33.
Safa A. Roles of c-FLIP in apoptosis, necroptosis, and autophagy. J Carcinog Mutagen. 2013;6:003. PubMed PMID: 25379355
Haider A, Duculan J, Whynot J, Krueger J. Increased JunB mRNA and protein expression in Psoriasis vulgaris lesions. J Invest Dermatol. 2006;126:912–4.
Imam A, Al Hasan M, El-Sayed S, Ibrahim S. The expression pattern of antiapoptotic protein c-FLIP in psoriatic epidermis. Egypt J Dermatol Venerol. 2013; 33:46-50.
Bagasra, O. Protocols for the in situ PCR-amplification and detection of mRNA and DNA sequences. Nat Protoc. 2007;2:2782-95.
Franklin R, Feng P, Milon B, Desouki M, Singh K, Kajdacsy-Balla A, et al. hZIP1 zinc uptake transporter down regulation and zinc depletion in prostate cancer. Mol Cancer. 2005;4:32. PubMed PMID: 16153295
Sinniah B, Saraswathy Devi S, Prashant B. Epidemiology of Psoriasis in Malaysia: a hospital-based study. Med J Malaysia. 2010;65:112-4.
Varghese F, Bukhari A, Malhotra R, De A. IHC profiler: An open-source plugin for the quantitative evaluation and automated scoring of immunohistochemistry images of human tissue samples. PLoS One. 2014;9:e96801. PubMed PMID: 24802416
Alexis A, Blackcloud P. Psoriasis in skin of color: epidemiology, genetics, clinical presentation, and treatment nuances. J Clin Aesthet Dermatol. 2014;7:16-24.
Huang R, Li L, Wang M, Chen X, Huang Q, Lu C. An exploration of the role of MicroRNAs in Psoriasis:A systematic review of the literature. Medicine. 2015;94:e2030. PubMed PMID: 26559308
Ransick A. Detection of mRNA by in situ hybridization and RT-PCR. Methods Cell Biol. 2004;74:601-20.
Xu N, Meisgen F, Butler L, Han G, Wang X, Söderberg-Nauclér C, et al. MicroRNA-31 is overexpressed in Psoriasis and modulates inflammatory cytokine and chemokine production in keratinocytes via targeting serine/threonine kinase 40. J Immunol. 2013;190:678-88.
Tsoi L, Spain S, Knight J, Ellinghaus E, Stuart P, Capon F, et al. Identification of 15 new psoriasis susceptibility loci highlights the role of innate immunity. Nat Genet. 2012;44:1341-8.
Nestle F, Kaplan D, Barker J. Psoriasis. N Engl J Med. 2009;361:496-509.
Kulski J, Kenworthy W, Bellgard M, Taplin R, Okamoto K, Oka A, et al. Gene expression profiling of Japanese psoriatic skin reveals an increased activity in molecular stress and immune response signals. J Mol Med. 2005;83:964-75.
Adamski Z, Kanabaj K, Kuźniak A. The link between psoriasis and other diseases based on epidemiological and genetic analyses. Postepy Dermatol Alergol. 2023;40:496-503.
Patel H, Revankar R, Pedroza S, Graham S, Feldman S. The genetic susceptibility to Psoriasis and the relationship of linked genes to our treatment options. Int J Mol Sci. 2023;24:12310. PubMed PMID: 37569685
Park C, Kim K, Woo S, Chun J, Lee K. Comparison of the expression profile of JunB, c-Jun, and S100A8 (calgranulin A) in Psoriasis vulgaris and guttate psoriasis. Ann Dermatol. 2009;21:35-8.
Kiafar B, Binabaj M, Jafarian A, Khazan Z, Hashemy S. The Relationship between tissue thioredoxin reductase activity and the psoriasis area and severity index. Indian J Dermatol. 2020;65:29-32.
Dilek N, Dilek A, Taşkın Y, Erkinüresin T, Yalçın Ö, Saral Y. Contribution of myeloperoxidase and inducible nitric oxide synthase to pathogenesis of psoriasis. Postepy Dermatol Alergol. 2016;33:435-9.
Yildirim M, Inaloz H, Baysal V, Delibas N. The role of oxidants and antioxidants in Psoriasis. J Eur Acad Dermatol Venereol. 2003;17:34-6.
El-Komy M, Amin I, El-Hawary M, Saadi D, Shaker O. Upregulation of the miRNA-155, miRNA-210, and miRNA-20b in psoriasis patients and their relation to IL-17. Int J Immunopathol Pharmacol. 2020;34: 2058738420933742. PubMed PMID: 32602388
