Department of Gynecology, WuHan Third Hospital (Tongren Hospital of WuHan University), Wuhan City, Hubei Province, China.
Department of Obstetrics and Gynecology, XiangYang First People's Hospitai Affiliated to Hubei Medicine, Xiangyang City, Hubei Province, China.
J Physiol Pharmacol. 2023 Dec;74(6). doi: 10.26402/jpp.2023.6.06. Epub 2024 Feb 7.
Paeonol (2-hydroxy-4-methoxyphenylacetophenone) is a natural phenolic component isolated from the root bark of peony with multiple pharmacological activities and has been proven to have anti-cancer effects. The objective of this study is to investigate the influence mechanism of paeonol on the proliferatory and apoptotic activities of ovarian cancer (OC) cells by modulating the transforming growth factor beta (TGF-β)/Smad3 pathway. The SKOV3 cells were pretreated with various concentrations of paeonol (0, 25, 50, 100, 200, 400 μg/mL) for 48 hours to determine the optimal experimental concentration of paeonol. Following this, the TGF-β overexpression vector was constructed and transfected into the SKOV3 cells. The assessment of cell proliferation, invasion, and migration was conducted through MTT, colony formation, flow cytometry, transwell, and wound-healing experiments. The detection of TGF-β/Smad3 pathway-related proteins and apoptosis-related proteins (B-cell lymphoma (Bcl-2) Bcl-2-associated X protein (Bax)) was performed using Western blot analysis. Paeonol exhibited a significant inhibitory effect on SKOV3 cell viability when administered at concentrations ranging from 50-400 μg/mL, with an IC value of 200 μg/mL. Within the concentration range of 50 to 200 μg/mL, paeonol exhibited a dose-dependent effect on the progression of SKOV3 cells, including a reduction in the anti-apoptotic protein Bcl-2, an increase in the pro-apoptotic protein Bax (P<0.05), inhibition of cell migration and invasion (P<0.05), and promotion of cell apoptosis (P<0.05), particularly at a concentration of 200 μg/mL. These effects were found to be more pronounced. The aforementioned effects of paeonol can be ascribed to its inhibition of the TGFβ/Smad3 pathway, according to a mechanistic viewpoint. It is noteworthy that the inhibitory impact of paeonol on SKOV3 cell progression is counteracted by the elevation of TGF-β levels following overexpression. We conclude that paeonol exerts regulatory effects on the TGF-β/Smad3 pathway, leading to the inhibition of proliferation, migration, and invasion of OC cells, thereby attenuating malignant behavior of cancer cells.
丹皮酚（2-羟基-4-甲氧基苯乙酮）是从牡丹根皮中分离得到的一种具有多种药理活性的天然酚类成分，已被证明具有抗癌作用。本研究旨在通过调节转化生长因子β（TGF-β）/Smad3 通路，探讨丹皮酚对卵巢癌细胞增殖和凋亡活性的影响机制。将 SKOV3 细胞用不同浓度的丹皮酚（0、25、50、100、200、400μg/ml）预处理 48 小时，以确定丹皮酚的最佳实验浓度。然后构建 TGF-β过表达载体并转染 SKOV3 细胞。通过 MTT、集落形成、流式细胞术、Transwell 和划痕愈合实验评估细胞增殖、侵袭和迁移。采用 Western blot 分析检测 TGF-β/Smad3 通路相关蛋白和凋亡相关蛋白（B 细胞淋巴瘤（Bcl-2）Bcl-2 相关 X 蛋白（Bax））。丹皮酚在 50-400μg/ml 浓度范围内对 SKOV3 细胞活力有显著的抑制作用，IC50 值为 200μg/ml。在 50-200μg/ml 浓度范围内，丹皮酚对 SKOV3 细胞的进展呈剂量依赖性，包括抗凋亡蛋白 Bcl-2 减少，促凋亡蛋白 Bax 增加（P<0.05），细胞迁移和侵袭抑制（P<0.05），促进细胞凋亡（P<0.05），特别是在 200μg/ml 浓度下作用更明显。从机制上看，这些作用可归因于丹皮酚对 TGFβ/Smad3 通路的抑制。值得注意的是，过表达后 TGF-β水平升高会抵消丹皮酚对 SKOV3 细胞进展的抑制作用。综上所述，丹皮酚对 TGF-β/Smad3 通路发挥调节作用，从而抑制 OC 细胞的增殖、迁移和侵袭，减弱癌细胞的恶性行为。
