Amar Saroj Kumar, Srivastav Ajeet K, Dubey Divya, Chopra Deepti, Singh Jyoti, Mujtaba Syed Faiz
1 Department of Forensic Science, School of Bioengineering and Biosciences, Lovely Professional University, Phagwara, Punjab, India.
2 Department of Biochemistry, School of Dental Sciences, Babu Banarasi Das University, Lucknow, Uttar Pradesh, India.
Toxicol Ind Health. 2019 Jul;35(7):457-465. doi: 10.1177/0748233719862128.
Solar ultraviolet (UV) radiation is the main factor of photocarcinogenesis, photoaging, and photosensitivity; thus protection from biological damaging UV radiation is a concern. Sunscreens containing UV filters are the most preferred means of photoprotection but the safety and efficacy of UV filters are in question. Benzophenone (BP) and its derivatives, namely, benzophenone 1 (BP1), is commonly used in sunscreens as a UV blocker. The aim of this study was to assess the effects of BP and BP1 on the differential expression of proteins in human keratinocytes (HaCaT cells) under exposure to ultraviolet A radiation. Photosensitive proteins were screened from HaCaT cells by two-dimensional (2-D) gel electrophoresis, and identification of these differentially expressed proteins was performed by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF)/TOF mass spectrometry. Protein identification was performed using the search program MASCOT and a database made of SUMO and GhJMJ12 amino acid sequences. Our results showed that the proteins involved directly or indirectly in apoptosis are 70 kDa heat shock protein, long-chain specific acyl-CoA dehydrogenase, serine/threonine-protein kinase, and FAM78A protein, which were upregulated in comparison to control HaCaT cells. The expressions of binding immunoglobulin protein, podocalyxin-like protein, actin, cytoplasmic, and calreticulin precursors were downregulated. The altered protein expression indicated that cell growth arrest and apoptosis were potential mechanisms of cytotoxicity and genotoxicity of BPs. The results of 2-D gel electrophoresis followed by mass spectrometry showed expression of novel proteins involved in promoting or initiating apoptotic pathways. Hence, we conclude that BPs should be avoided as a UV blocker from sunscreens because of its potential to promote apoptotic proteins in human skin keratinocytes.
太阳紫外线(UV)辐射是光致癌、光老化和光过敏的主要因素;因此,防止生物性损伤的紫外线辐射是一个值得关注的问题。含有紫外线过滤剂的防晒霜是最常用的光防护手段,但紫外线过滤剂的安全性和有效性存在疑问。二苯甲酮(BP)及其衍生物,即二苯甲酮1(BP1),常用于防晒霜中作为紫外线阻滞剂。本研究的目的是评估BP和BP1在紫外线A辐射暴露下对人角质形成细胞(HaCaT细胞)中蛋白质差异表达的影响。通过二维(2-D)凝胶电泳从HaCaT细胞中筛选光敏蛋白,并通过基质辅助激光解吸/电离飞行时间(MALDI-TOF)/TOF质谱对这些差异表达的蛋白质进行鉴定。使用搜索程序MASCOT和由SUMO和GhJMJ12氨基酸序列组成的数据库进行蛋白质鉴定。我们的结果表明,与对照HaCaT细胞相比,直接或间接参与细胞凋亡的蛋白质有70 kDa热休克蛋白、长链特异性酰基辅酶A脱氢酶、丝氨酸/苏氨酸蛋白激酶和FAM78A蛋白,它们的表达上调。结合免疫球蛋白蛋白、足突蛋白样蛋白、肌动蛋白、细胞质和钙网蛋白前体的表达下调。蛋白质表达的改变表明细胞生长停滞和细胞凋亡是二苯甲酮细胞毒性和遗传毒性的潜在机制。二维凝胶电泳随后进行质谱分析的结果显示了参与促进或启动凋亡途径的新蛋白质的表达。因此,我们得出结论,由于二苯甲酮有可能促进人皮肤角质形成细胞中的凋亡蛋白,应避免将其作为防晒霜中的紫外线阻滞剂。