Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland.
J Cell Physiol. 2014 Aug;229(8):1028-38. doi: 10.1002/jcp.24527.
Stearoyl-CoA desaturase (SCD, SCD1), an endoplasmic reticulum (ER) resident protein and a rate-limiting enzyme in monounsaturated fatty acid biosynthesis, regulates cellular functions by controlling the ratio of saturated to monounsaturated fatty acids. Increase in SCD expression is strongly implicated in the proliferation and survival of cancer cells, whereas its decrease is known to impair proliferation, induce apoptosis, and restore insulin sensitivity. We examined whether fenretinide, (N-(4-hydroxyphenyl)retinamide, 4HPR), which induces apoptosis in cancer cells and recently shown to improve insulin sensitivity, can modulate the expression of SCD. We observed that fenretinide decreased SCD protein and enzymatic activity in the ARPE-19 human retinal pigment epithelial cell line. Increased expression of BiP/GRP78, ATF4, and GADD153 implicated ER stress. Tunicamycin and thapsigargin, compounds known to induce ER stress, also decreased the SCD protein. This decrease was completely blocked by the proteasome inhibitor MG132. In addition, PYR41, an inhibitor of ubiquitin activating enzyme E1, blocked the fenretinide-mediated decrease in SCD. Immunoprecipitation analysis using anti-ubiquitin and anti-SCD antibodies and the blocking of SCD loss by PYR41 inhibition of ubiquitination further corroborate that fenretinide mediates the degradation of SCD in human RPE cells via the ubiquitin-proteasome dependent pathway. Therefore, the effect of fenretinide on SCD should be considered in its potential therapeutic role against cancer, type-2 diabetes, and retinal diseases.
硬脂酰辅酶 A 去饱和酶(SCD,SCD1)是内质网(ER)驻留蛋白,也是单不饱和脂肪酸生物合成的限速酶,通过控制饱和脂肪酸与单不饱和脂肪酸的比例来调节细胞功能。SCD 表达的增加强烈暗示了癌细胞的增殖和存活,而其减少已知会损害增殖、诱导细胞凋亡,并恢复胰岛素敏感性。我们研究了 fenretinide(N-(4-羟苯基)视黄酰胺,4HPR)是否可以调节 SCD 的表达,fenretinide 可诱导癌细胞凋亡,并最近显示出改善胰岛素敏感性的作用。我们观察到 fenretinide 降低了人视网膜色素上皮细胞系 ARPE-19 中的 SCD 蛋白和酶活性。BiP/GRP78、ATF4 和 GADD153 的表达增加表明内质网应激。已知诱导内质网应激的化合物 tunicamycin 和 thapsigargin 也降低了 SCD 蛋白。这种减少被蛋白酶体抑制剂 MG132 完全阻断。此外,泛素激活酶 E1 的抑制剂 PYR41 阻断了 fenretinide 介导的 SCD 减少。使用抗泛素和抗 SCD 抗体进行免疫沉淀分析以及 PYR41 抑制泛素化对 fenretinide 介导的 SCD 丢失的阻断进一步证实,fenretinide 通过泛素-蛋白酶体依赖性途径介导人 RPE 细胞中 SCD 的降解。因此,在考虑 fenretinide 治疗癌症、2 型糖尿病和视网膜疾病的潜在治疗作用时,应考虑其对 SCD 的影响。