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      Ultraviolet B (UVB) Irradiation-Induced Apoptosis in Various Cell Lineages in Vitro

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          Abstract

          Ultraviolet B (UVB) radiation acts as a strong apoptotic trigger in many cell types, in tumor and normal cells. Several studies have demonstrated that UVB-induced cell death occurs through the generation of reactive oxygen species. The consequent oxidative stress includes the impairment of cellular antioxidants, the induction of DNA damage and the occurrence of apoptosis. In this review, we investigated UVB apoptotic action in various cell models by using ultrastructural, molecular and cytofluorimetric techniques. Myeloid leukemia HL-60, T-lymphoblastoid Molt-4 and myelomonocytic U937 human cells, generally affected by apoptotic stimuli, were studied. Human chondrocytes and C2C12 skeletal muscle cells, known to be more resistant to damage, were also considered. All of them, when exposed to UVB radiation, revealed a number of characteristic apoptotic markers. Membrane blebbing, cytoplasm shrinkage and chromatin condensation were detected by means of electron microscopy. DNA cleavage, investigated by using agarose gel electrophoresis and TUNEL reaction, was observed in suspended cells. Differently, in chondrocytes and in skeletal muscle cells, oligonucleosomic DNA fragmentation did not appear, even if a certain TUNEL positivity was detected. These findings demonstrate that UVB radiation appears to be an ideal tool to study the apoptotic behavior.

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          C2C12 murine myoblasts as a model of skeletal muscle development: morpho-functional characterization.

          In this study, the differentiation of C2C12 cells, a primary line of murine myoblasts, was investigated by a multiple technical approach. Undifferentiated cells, and those at intermediate and final differentiation times, were studied at the reverted microscope, by conventional and confocal immunofluorescence, and by transmission and scanning electron microscopy. The general monolayer architecture changed during differentiation from fusiform or star-shaped cells to elongated confluent cells, finally originating long, multinucleated myotubes. Sarcomeric actin and myosin are present also in undifferentiated myoblasts, but progressively acquire a structured pattern up to the appearance of sarcomeres and myofibrils at about 5 days after differentiation induction. Myotubes show a particular positivity for actin and myosin, and M-cadherin, an adhesion molecule characteristic, as known, of satellite cells, also seems to be involved in their assembling. Rare apoptotic patterns, as evidenced by the TUNEL technique, appear during myoblast maturation.
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            Light, including ultraviolet.

            Ultraviolet (UV) light is intricately linked to the functional status of the cutaneous immune system. In susceptible individuals, UV radiation can ignite pathogenic inflammatory pathways leading to allergy or autoimmunity. In others, this same UV radiation can be used as a phototherapy to suppress pathogenic cutaneous immune responses. These vastly different properties are a direct result of UV light's ability to ionize molecules in the skin and thereby chemically alter them. Sometimes these UV-induced chemical reactions are essential, the formation of pre-vitamin D(3) from 7-dehydrocholesterol, for example. In other instances they can be potentially detrimental. UV radiation can ionize a cell's DNA causing adjacent pyrimidine bases to chemically bond to each other. To prevent malignant transformation, a cell may respond to this UV-induced DNA damage by undergoing apoptosis. Although this pathway prevents skin cancer it also has the potential of inducing or exacerbating autoreactive immune responses by exposing the cell's nuclear antigens. Ultraviolet-induced chemical reactions can activate the immune system by a variety of other mechanisms as well. In response to UV irradiation keratinocytes secrete cytokines and chemokines, which activate and recruit leukocytes to the skin. In some individuals UV-induced chemical reactions can synthesize novel antigens resulting in a photoallergy. Alternatively, photosensitizing molecules can damage cells by initiating sunburn-like phototoxic reactions. Herein we review all types of UV-induced skin reactions, especially those involving the immune system. Published by Elsevier Ltd.
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              UVB-induced DNA damage, generation of reactive oxygen species, and inflammation are effectively attenuated by the flavonoid luteolin in vitro and in vivo.

              Ultraviolet (UV) radiation induces DNA damage, oxidative stress, and inflammatory processes in human keratinocytes resulting in skin inflammation, photoaging, and photocarcinogenesis. The flavonoid luteolin is one of the most potent antioxidative plant polyphenols. We investigated the UV protective and antioxidant properties of luteolin in human keratinocytes in vitro, ex vivo, and in vivo. Spectrophotometric measurements revealed extinction maxima of luteolin in the UVB and UVA range. UV transmission below 370 nm was <10%. In human skin, luteolin effectively reduced the formation of UVB-induced cyclobutane pyrimidine dimers. The free radical scavenging activity of luteolin was assessed in various cell-free and cell-based assays. In the cell-free DPPH assay the half-maximal effective concentration (EC₅₀) of luteolin (12 μg/ml) was comparable to those of Trolox (25 μg/ml) and N-acetylcysteine (32 μg/ml). In contrast, in the H₂DCFDA assay performed with UVB-irradiated keratinocytes, luteolin (EC₅₀ 3 μg/ml) was much more effective compared to Trolox (EC₅₀ 12 μg/ml) and N-acetylcysteine (EC₅₀ 847 μg/ml). Luteolin also inhibited both UVB-induced skin erythema and the upregulation of cyclooxygenase-2 and prostaglandin E₂ production in human skin via interference with the MAPK pathway. These data suggest that luteolin may protect human skin from UVB-induced damage by a combination of UV-absorbing, DNA-protective, antioxidant, and anti-inflammatory properties. Copyright © 2011 Elsevier Inc. All rights reserved.
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                Author and article information

                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                January 2013
                27 December 2012
                : 14
                : 1
                : 532-546
                Affiliations
                [1 ]DiSTeVA, University of Urbino “Carlo Bo”, Urbino 61029, Italy; E-Mails: sara.salucci@ 123456uniurb.it (S.S.); sabrina.burattini@ 123456uniurb.it (S.B.); michela.battistelli@ 123456uniurb.it (M.B.); valentina.baldassarri@ 123456uniurb.it (V.B.)
                [2 ]Laboratory of Musculoskeletal Cell Biology, Rizzoli Orthopaedic Institute, Bologna 40136, Italy; E-Mail: mariacristina.maltarello@ 123456ior.it
                [3 ]IGM, CNR, Rizzoli Orthopaedic Institute, Bologna 40136, Italy
                Author notes
                [* ]Author to whom correspondence should be addressed; E-Mail: elisabetta.falcieri@ 123456uniurb.it ; Tel.: +39-0722304284; Fax: +39-0722304244.
                Article
                ijms-14-00532
                10.3390/ijms14010532
                3565280
                23271369
                b3a82c43-9214-49a2-9f08-0bb6ac49a174
                © 2013 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

                History
                : 31 October 2012
                : 19 December 2012
                : 21 December 2012
                Categories
                Review

                Molecular biology
                uvb,apoptosis,hl-60,molt-4,u937,chondrocytes,skeletal muscle cells
                Molecular biology
                uvb, apoptosis, hl-60, molt-4, u937, chondrocytes, skeletal muscle cells

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