Mechanisms of Chemically Induced Photosensitivity

化学诱导光敏性的机制

基本信息

批准号：
7593921
负责人：
COLIN CHIGNELL
金额：
$ 146.07万
依托单位：
NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7593921
关键词：
Alkaloids Amphiregulin Attenuated Berberine Biological Assay Blocking Antibodies Cell Cycle Progression Cell Proliferation Cells Chemopreventive Agent Cyclin D1 Development Dominant-Negative Mutation Dose Environmental Hazards Epidermal Growth Factor Receptor Flame Retardants GM 6001 Goldenseal Human Hypericin Hypericum perforatum Incubated Light Lipids Maps Mediating Metalloproteases Mitogen-Activated Protein Kinase Inhibitor Molecular Oxygen PI3K/AKT Patients Photosensitivity Photosensitizing Agents Plasma Process Proto-Oncogene Proteins c-akt Research Project Grants Role Signal Pathway Singlet Oxygen Skin Small Interfering RNA Smith-Lemli-Opitz Syndrome Structure of retinal pigment epithelium Superoxides Techniques Therapeutic Therapeutic Effect Triethylenetetramine UVA induced Visible Radiation inhibitor/antagonist interest irradiation keratinocyte kinase inhibitor lens oxidation tetrabromobisphenol A

项目摘要

We have demonstrated that a low, non-lethal dose of UVA exposure induces dose-dependent cell cycle progression in human HaCaT keratinocytes. We found that UVA induced cyclin D1 accumulation, while siRNA knockdown of cyclin D1 blocked the UVA-induced cell cycle progression, indicating that this process is mediated by cyclin D1. UVA irradiation also induced AKT activation; when cells were incubated with PI3K/AKT inhibitor or infected with dominant negative AKT, cell cycle progression and proliferation were inhibited, suggesting that AKT activation is involved in UVA-induced cell cycle progression. In contrast, ERK was not activated by UVA exposure; incubation with ERK/MAPK inhibitor had no effect on UVA-induced cell cycle progression. Activation of EGFR was observed after UVA exposure. EGFR kinase inhibitor AG1478 attenuated UVA-induced cell cycle progression, indicating the involvement of EGFR activation. Furthermore, metalloprotease inhibitor GM6001 blocked UVA-induced cell cycle progression, and siRNA knockdown of ADAM17 had a similar inhibitory effect, demonstrating that ADAM17 has a role in mediating the UVA-induced cell cycle progression. Consistent with the role of ADAM17, antibody blocking amphiregulin attenuated UVA-induced cell cycle progression, implying the involvement of amphiregulin. Identification of these signaling pathways in UVA-induced cell proliferation will facilitate the development of efficient and safe chemopreventive and therapeutic strategies for skin cancer.We have identified the skin lipid cholesta-5,7,9(11)-trien-3 beta-ol (9-DDHC) as the putative agent responsible for UVA-induced skin photosensitivity in Smith-Lemli-Opitz syndrome patients. 9-DDHC generates singlet oxygen and superoxide upon UVA irradiation, is phototoxic to keratinocytes and is found in higher concentrations in plasma from UVA sensitive patients. We have demonstrated that berberine, the main alkaloid present in Goldenseal, is phototoxic to human retinal pigment epithelial cells and lens cells, while hypericin, found in St Johns Wort, is photototoxic to retinal pigment epithelial cells. We have successfully mapped the intracellular spatial localization of singlet oxygen in keratinocytes using the immuno-spin assay technique. Finally, We have found that the flame retardant 3,3,5,5-tetrabromobisphenol A (TBBPA) is subject to photosensitized oxidation involving singlet molecular oxygen. Wide use of flame retardants such as TBBPA can pose an environmental hazard and it is of interest to investigate how they may degrade.

我们已经证明，低剂量的UVA暴露会诱导人类HACAT角质形成细胞的剂量依赖性细胞周期进程。我们发现UVA诱导细胞周期蛋白D1的积累，而Cyclin D1的siRNA敲低阻断了UVA诱导的细胞周期进程，表明该过程是由Cyclin d1介导的。 UVA辐照也诱导Akt激活。当细胞与PI3K/AKT抑制剂孵育或感染显性阴性AKT时，细胞周期进程和增殖被抑制，这表明AKT激活与UVA诱导的细胞周期进程有关。相比之下，ERK没有被UVA暴露激活。与ERK/MAPK抑制剂一起孵育对UVA诱导的细胞周期进程没有影响。 UVA暴露后观察到EGFR的激活。 EGFR激酶抑制剂AG1478减弱了UVA诱导的细胞周期进程，表明EGFR激活的参与。此外，金属蛋白酶抑制剂GM6001阻断了UVA诱导的细胞周期进程，ADAM17的siRNA敲低具有相似的抑制作用，表明ADAM17在介导UVA诱导的细胞周期进程中起作用。与ADAM17的作用一致，抗体阻断两极分裂蛋白会减弱UVA诱导的细胞周期进程，这意味着两极分裂蛋白的参与。 Identification of these signaling pathways in UVA-induced cell proliferation will facilitate the development of efficient and safe chemopreventive and therapeutic strategies for skin cancer.We have identified the skin lipid cholesta-5,7,9(11)-trien-3 beta-ol (9-DDHC) as the putative agent responsible for UVA-induced skin photosensitivity in Smith-Lemli-Opitz syndrome patients. 9-DDHC在UVA辐照时会产生单线氧和超氧化物，对角质形成细胞具有光毒性，并且在UVA敏感患者的血浆中发现了较高浓度。我们已经证明，黄晶是金素中存在的主要生物碱，是对人视网膜色素上皮细胞和晶状体细胞的光毒性，而在圣约翰斯豆类中发现的蛋白质是对视网膜色素上皮细胞的光毒素。我们已经成功地映射了使用免疫旋转测定技术在角质形成细胞中单线氧的细胞内空间定位。最后，我们发现阻燃剂3,3,5,5-四甲醇A（TBBPA）受到涉及单链分子氧的光敏氧化。大量使用阻燃剂（例如TBBPA）可能构成环境危害，并且有趣的是研究它们如何降解。