Department of Anesthesiology, The First Hospital of Jilin University, Changchun, Jilin, China (mainland).
Department of Hand and Foot Surgery, The First Hospital of Jilin University, Changchun, Jilin, China (mainland).
Med Sci Monit. 2018 Jan 25;24:486-502. doi: 10.12659/msm.906049.
BACKGROUND A recent focus in skin cancer prevention intervenes though modulating molecular links between inflammation and cell growth signaling, such as NF-κB. This study elucidates the effect of a non-tumor promoting phorbol ester, ingenol-3-angelate (I3A), on the growth of human melanoma cells and on the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin inflammation and 7,12-Dimethylbenz(a)anthracene (DMBA)-induced skin carcinoma in mice. MATERIAL AND METHODS Cell viability was assessed by MTT assay, cell proliferation by clonogenic assay, apoptosis and cell cycle arrest was analyzed by flow cytometry, protein expression was studied by IHC and Western blotting, and gene expression by qPCR. RESULTS I3A suppressed the survival and proliferation of human melanoma cells with estimated IC50 values around 38 and 46 μM for A2058 and HT144 cell, respectively. I3A activated the protein levels of PKCδ and PKCε, which induced apoptosis by activating caspase-9 and caspace-3 followed by lowering of mitochondrial membrane potential and enhancing DNA fragmentation. I3A induced G1 phase cell cycle arrest as well as G2/M phase arrest in both cell lines. I3A inhibited the levels of NFκB p65 protein as well as phosphorylation of p65 and its nuclear translocation. I3A suppressed the gene expression of NF-κB, COX-2 and iNOS. I3A inhibited TPA-induced inflammation and epidermal hyperplasia in female ICR mice by downregulating NF-κB and iNOS. I3A suppressed the growth of skin tumor in DMBA-induced mice in dose-dependent manner. CONCLUSIONS The mechanism of I3A induces apoptosis in human melanoma cells and suppresses skin inflammation and carcinoma via downregulation of NF-κB-iNOS-COX-2 signaling.
最近的皮肤癌预防研究重点是通过调节炎症和细胞生长信号之间的分子联系，如 NF-κB，来干预皮肤癌。本研究阐明了非促肿瘤佛波醇酯——表没食子儿茶素没食子酸酯（I3A）对人黑色素瘤细胞生长的影响，以及对 12-O-十四烷酰佛波醇-13-乙酸酯（TPA）诱导的皮肤炎症和 7,12-二甲基苯并蒽（DMBA）诱导的皮肤癌的影响。
通过 MTT 法评估细胞活力，通过集落形成试验评估细胞增殖，通过流式细胞术分析细胞凋亡和细胞周期停滞，通过免疫组化和 Western blot 研究蛋白质表达，通过 qPCR 研究基因表达。
I3A 抑制了人黑色素瘤细胞的存活和增殖，A2058 和 HT144 细胞的 IC50 值分别约为 38 和 46 μM。I3A 激活了 PKCδ 和 PKCε 的蛋白水平，通过激活 caspase-9 和 caspase-3 诱导凋亡，随后降低线粒体膜电位并增强 DNA 片段化。I3A 在两种细胞系中均诱导 G1 期细胞周期停滞和 G2/M 期停滞。I3A 抑制了 NFκB p65 蛋白及其磷酸化和核转位的水平。I3A 抑制了 NF-κB、COX-2 和 iNOS 的基因表达。I3A 通过下调 NF-κB 和 iNOS 抑制了 TPA 诱导的雌性 ICR 小鼠的炎症和表皮增生。I3A 以剂量依赖的方式抑制 DMBA 诱导的小鼠皮肤肿瘤的生长。
I3A 通过下调 NF-κB-iNOS-COX-2 信号通路诱导人黑色素瘤细胞凋亡，并抑制皮肤炎症和癌。
