Institute of Immunology, Faculty of Medicine, Central University of Venezuela, Caracas, Venezuela.
Biotecnology Unit, Faculty of Pharmacy, Central University of Venezuela, Caracas, Venezuela.
Anticancer Agents Med Chem. 2023;23(10):1145-1155. doi: 10.2174/1871520622666220415105615.
Kaurane-type diterpenoids, obtained from various natural sources, have shown many biological activities, including anti-inflammatory and antitumor effects. Caracasine, an -kaurane diterpenoid isolated from the flowers of , was shown to induce apoptosis in leukaemia cell lines.
The present study aimed to ascertain the compound's mechanism of cell death induction using two leukaemia cell lines, Jurkat E6.1 (T cell) and HL-60 (promyeloblast cells).
Cell death in Jurkat and HL60 cells were evaluated by flow cytometry for apoptosis with annexin-V/PI, mitochondrial membrane potential disturbance, changes in cell cycle, CD95 expression, caspase activation, Nuclear Factor kappa B inhibition, and differentiation into a neutrophil-like cell (dHL60).
Caracasine (10 μM) increased the G0/G1 phase in Jurkat and arrested the cell cycle in the S phase in HL60. Caracasine increased CD95 expression (<0.01 in Jurkat and <0.05 in HL60) and caspase-8 activation (<0.001 in Jurkat and p<0.05 in HL60). Caspase-9 was activated in both cell lines (<0.001) along with the decline in mitochondrial Δψm (<0.05 in Jurkat and p<0.001 in HL60). In HL60 cells, the kaurane induced neutrophil differentiation was assessed by CD40 expression and reactive oxygen species production. In Jurkat cells, caracasine inhibited the NF-κB pathway in cells pretreated with PHA to activate the NF-κB pathway, suggesting a possible role in inflammatory diseases.
Caracasine induced apoptosis through the intrinsic and extrinsic pathways in both cell lines were evaluated which could be the leading structure for new anti-leukemic and anti-inflammatory drugs.
来源于各种天然来源的贝壳杉烷型二萜类化合物具有许多生物活性，包括抗炎和抗肿瘤作用。从 Caracas 花中分离得到的贝壳杉烷二萜 caracasine 已被证明可诱导白血病细胞系凋亡。
本研究旨在使用两种白血病细胞系 Jurkat E6.1（T 细胞）和 HL-60（早幼粒细胞）确定该化合物诱导细胞死亡的机制。
通过流式细胞术用 annexin-V/PI 评估 Jurkat 和 HL60 细胞中的细胞死亡情况，检测线粒体膜电位紊乱、细胞周期变化、CD95 表达、半胱天冬酶激活、核因子 kappa B 抑制和向中性粒细胞样细胞（dHL60）分化。
Caracasine（10 μM）增加 Jurkat 的 G0/G1 期，并使 HL60 的细胞周期停滞在 S 期。Caracasine 增加 CD95 表达（Jurkat 中<0.01，HL60 中<0.05）和半胱天冬酶-8 激活（Jurkat 中<0.001，HL60 中 p<0.05）。两种细胞系中 caspase-9 均被激活（<0.001），同时线粒体 Δψm 下降（Jurkat 中<0.05，HL60 中 p<0.001）。在 HL60 细胞中，通过 CD40 表达和活性氧物质产生评估贝壳杉烷诱导的中性粒细胞分化。在 Jurkat 细胞中，caracasine 抑制 PHA 预处理激活 NF-κB 通路的细胞中的 NF-κB 通路，提示其在炎症性疾病中可能具有作用。
Caracasine 通过两种细胞系中的内在和外在途径诱导凋亡，这可能是新型抗白血病和抗炎药物的先导结构。
