Department of Gynecology, Ningbo First Hospital, Ningbo, Zhejiang, China.
Department of Medical Laboratory, Ningbo First Hospital, Ningbo, Zhejiang, China.
Chem Biol Drug Des. 2023 Jul;102(1):101-114. doi: 10.1111/cbdd.14228. Epub 2023 Mar 14.
Evodiamine (EVO) has been demonstrated to promote apoptosis of ovarian cancer cells, and upregulate miR-152-3p level in colorectal cancer. Here, we explore part of the network mechanism of EVO and miR-152-3p in ovarian cancer. The bioinformatics website, dual luciferase reporter assay, and quantitative real-time polymerase chain reaction were applied to analyze the network among EVO, lncRNA, miR-152-3p, and mRNA. The effect and mechanism of EVO on ovarian cancer cells were determined using cell counting kit-8, flow cytometry, TUNEL, Western blot, and rescue experiments. As a result, EVO dose-dependently attenuated cell viability, induced G2/M phase arrest and apoptosis, promoted miR-152-3p level (4.5- or 2-fold changes), and inhibited expressions of NEAT1 (0.225- or 0.367-fold changes), CDK8 (0.625- or 0.571-fold changes), and CDK19 (0.25- or 0.147-fold changes) in OVCAR-3 and SKOV-3 cells. In addition, EVO decreased Bcl-2 expression, but increased the expressions of Bax and c-caspase-3. NEAT1 targeted miR-152-3p which bound to CDK19. The impacts of EVO on cell viability, cycle, apoptosis, and apoptosis-related proteins were partially reversed by miR-152-3p inhibitor, NEAT1 overexpression, or CDK19 overexpression. Furthermore, miR-152-3p mimic offset the effects of NEAT1 or CDK19 overexpression. The role of NEAT1 overexpression in the biological phenotype of ovarian cancer cells was counteracted by shCDK19. In conclusion, EVO attenuates ovarian cancer cell progression via the NEAT1-miR-152-3p-CDK19 axis.
吴茱萸碱（EVO）已被证明可促进卵巢癌细胞凋亡，并上调结直肠癌中的 miR-152-3p 水平。在这里，我们探索了 EVO 和 miR-152-3p 在卵巢癌中的部分网络机制。我们应用生物信息学网站、双荧光素酶报告基因检测和实时定量聚合酶链反应来分析 EVO、lncRNA、miR-152-3p 和 mRNA 之间的网络。使用细胞计数试剂盒-8、流式细胞术、TUNEL、Western blot 和挽救实验来确定 EVO 对卵巢癌细胞的影响和作用机制。结果显示，EVO 呈剂量依赖性地减弱细胞活力，诱导 G2/M 期阻滞和细胞凋亡，促进 miR-152-3p 水平（增加 4.5-或 2 倍），并抑制 NEAT1（减少 0.225-或 0.367 倍）、CDK8（减少 0.625-或 0.571 倍）和 CDK19（减少 0.25-或 0.147 倍）的表达在 OVCAR-3 和 SKOV-3 细胞中。此外，EVO 降低了 Bcl-2 的表达，但增加了 Bax 和 c-caspase-3 的表达。NEAT1 靶向 miR-152-3p，后者与 CDK19 结合。EVO 对细胞活力、周期、凋亡和凋亡相关蛋白的影响部分被 miR-152-3p 抑制剂、NEAT1 过表达或 CDK19 过表达所逆转。此外，miR-152-3p 模拟物抵消了 NEAT1 或 CDK19 过表达的作用。NEAT1 过表达在卵巢癌细胞生物学表型中的作用被 shCDK19 抵消。总之，EVO 通过 NEAT1-miR-152-3p-CDK19 轴来减弱卵巢癌细胞的进展。
