Colombero Cecilia, Papademetrio Daniela, Sacca Paula, Mormandi Eduardo, Alvarez Elida, Nowicki Susana
Centro de Investigaciones Endocrinológicas "Dr. César Bergadá" (CEDIE), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Fundación de Endocrinología Infantil, División de Endocrinología, Hospital de Niños Ricardo Gutiérrez, Gallo 1360, C1425EFD, Buenos Aires, Argentina.
Universidad de Buenos Aires, Facultad de Farmacia y Bioquimica - Instituto de Estudios de la Inmunidad Humoral Prof Ricardo Margni. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Junin 954, C1113AAD, Buenos Aires, Argentina.
Horm Cancer. 2017 Aug;8(4):243-256. doi: 10.1007/s12672-017-0299-0. Epub 2017 Jun 21.
20-Hydroxyeicosatetraenoic acid (20-HETE) is generated intracellularly through the ω-hydroxylation of arachidonic acid by the cytochrome P450 (in humans, CYP4A11 and CYP4F2). 20-HETE induces mitogenic responses in different cancer cells. The aim of this study was to analyze how 20-HETE impacts cell survival, proliferation, and apoptosis in prostate cancer cells. Incubation of the human androgen-sensitive cells (LNCaP) with 1-10 μM HET0016 (a selective inhibitor of 20-HETE synthesis) reduced cell viability by 49*-64%* (p < 0.05 vs. control). This was explained by a reduction in cell proliferation (vehicle, 46 ± 3%; 1 μM, 23 ± 3%; 10 μM, 28 ± 3%) and by an increase in apoptosis (vehicle, 2.1 ± 0%; 1 μM, 16 ± 4%; 10 μM, 31 ± 3%). Furthermore, the increase in LNCaP cell viability induced by dihydrotestosterone (DHT, 0.1 nM) was abrogated by 30-42%* by 1-10 μM HET0016. Incubation with 20-HETE (5-1000 nM) increased LNCaP cell viability up to 50%, together with a 70% reduction in apoptosis. PC-3 (androgen-insensitive) cell viability was not affected by either HET0016 or 20-HETE. In LNCaP cells, HET0016 (10 μM) diminished the expression of androgen receptors (AR): messenger RNA (mRNA) (40%) and protein (50%). DHT (10 nM) augmented CYP4F2 protein expression (1.9-fold*) and 20-HETE levels (50%*). Oppositely, enzalutamide (AR antagonist) reduced CYP4F2 mRNA and protein expressions by 30 and 25%, respectively. Thus, intracellular availability of 20-HETE is necessary to sustain LNCaP cell viability. 20-HETE may act as a signaling molecule in the pathways involved in LNCaP cell viability upon stimulation of the AR. This effect may be partially attributed to its role on securing normal AR expression levels that in turn contribute to maintain intracellular levels of 20-HETE.
20-羟基二十碳四烯酸（20-HETE）在细胞内由细胞色素P450（在人类中为CYP4A11和CYP4F2）将花生四烯酸ω-羟基化产生。20-HETE可诱导不同癌细胞的促有丝分裂反应。本研究的目的是分析20-HETE如何影响前列腺癌细胞的存活、增殖和凋亡。用1-10μM HET0016（20-HETE合成的选择性抑制剂）孵育人雄激素敏感细胞（LNCaP）可使细胞活力降低49%-64%（与对照组相比，p<0.05）。这可通过细胞增殖减少（溶剂对照组，46±3%；1μM，23±3%；10μM，28±3%）和凋亡增加（溶剂对照组，2.1±0%；1μM，16±4%；10μM，31±3%）来解释。此外，1-10μM HET0016可使二氢睾酮（DHT，0.1 nM）诱导的LNCaP细胞活力增加被消除30%-42%。用20-HETE（5-1000 nM）孵育可使LNCaP细胞活力增加高达50%，同时凋亡减少70%。PC-3（雄激素不敏感）细胞活力不受HET0016或20-HETE的影响。在LNCaP细胞中，HET0016（10μM）可使雄激素受体（AR）的表达降低：信使核糖核酸（mRNA）（40%）和蛋白质（50%）。DHT（10 nM）可使CYP4F2蛋白表达增加（1.9倍*）和20-HETE水平增加（50%*）。相反，恩杂鲁胺（AR拮抗剂）可使CYP4F2 mRNA和蛋白表达分别降低30%和25%。因此，20-HETE的细胞内可用性对于维持LNCaP细胞活力是必要的。20-HETE可能在AR刺激后参与LNCaP细胞活力的信号通路中作为信号分子发挥作用。这种作用可能部分归因于其在确保正常AR表达水平方面的作用，而这反过来又有助于维持细胞内20-HETE的水平。
