2025 年第 4 期 第 20 卷

松果菊苷对卵巢癌细胞生物学行为和血管生成的影响及机制研究

Study on the effect and mechanism of echinacoside on the biological behavior and angiogenesis of ovarian cancer cells

作者：苏园1焦阳1夏新凤2张娜3

英文作者：Su Yuan1 Jiao Yang1 Xia Xinfeng2 Zhang Na3

单位：1南京中医药大学附属南京医院江苏省南京市第二医院药学部，南京210003；2江苏省徐州市睢宁县人民医院临床药学科，徐州221225；3江苏省中医院药剂科，南京210029

英文单位：1Department of Pharmacy the Second Hospital of Nanjing Jiangsu Province Nanjing Hospital Affiliated to Nanjing University of Chinese Medicine Nanjing 210003 China; 2Department of Clinical Pharmacy Suining County People′s Hospital of Xuzhou City Jiangsu Procince Xuzhou 221225 China; 3Department of Pharmacy Jiangsu Province Hospital of Chinese Medicine Nanjing 210029 China

关键词：卵巢癌；松果菊苷；恶性进展；磷脂酰肌醇-3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路

英文关键词：Ovariancancer;Echinacoside;Malignantprogression;Phosphatidylinositol-3-kinase/proteinkinaseB/mammaliantargetofrapamycinsignalingpathway

• 摘要：

• 目的 探讨松果菊苷对卵巢癌细胞增殖、迁移、侵袭和血管生成的影响及其可能机制。方法 以卵巢浆液性癌细胞株SKOV3为研究对象，细胞计数盒8法检测不同浓度松果菊苷（0、6.25、12.5、25、50、100、200、400 μmol/L）对细胞活力的影响；将SKOV3细胞完全随机分为对照组、松果菊苷低剂量（25 μmol/L）组、松果菊苷中剂量（50 μmol/L）组、松果菊苷高剂量（100 μmol/L）组和松果菊苷高剂量+740Y-P［磷脂酰肌醇-3-激酶（PI3K）激动剂，100 μmol/L］组。采用集落形成实验、TUNEL染色和Transwell实验检测细胞增殖、凋亡、迁移和侵袭能力；成管试验检测血管生成；蛋白质印迹法检测增殖、凋亡、侵袭、血管生成和PI3K/蛋白激酶B（AKT）/哺乳动物雷帕霉素靶蛋白（mTOR）通路相关蛋白表达。结果 细胞计数盒8法检测结果发现，随着松果菊苷浓度的增加，SKOV3细胞的存活率逐渐降低，分别为（100.0±5.5）%、（98.3±3.3）%、（93.3±3.4）%、（58.3±3.8）%、（42.4±4.6）%、（28.8±2.2）%、（19.4±3.4）%、（6.4±1.5）%，呈浓度依赖性（P<0.05）。与对照组比较，松果菊苷低、中和高剂量组SKOV3细胞的集落形成数、迁移和侵袭细胞数、相对环形管状结构形成比、细胞增殖抗原Ki-67、B淋巴细胞瘤基因2（Bcl-2）、N-钙黏蛋白（N-cadherin）和血管内皮生长因子A（VEGFA）表达，磷酸化（p）-PI3K/PI3K、p-AKT（S473）/AKT、p-AKT（T308）/AKT和p-mTOR/mTOR比值均逐渐减少/降低，凋亡率和活化半胱氨酸蛋白酶3（Cleaved caspase 3）、Bcl-2相关X蛋白（Bax）、E-钙黏蛋白（E-cadherin）表达逐渐升高，呈浓度依赖性，差异均有统计学意义（均P<0.05）。与松果菊苷高剂量组比较，松果菊苷高剂量+740Y-P组SKOV3细胞的集落形成数、迁移和侵袭细胞数、相对环形管状结构形成比，Ki-67、Bcl-2、N-cadherin和VEGFA表达，p-PI3K/PI3K、p-AKT（S473）/AKT、p-AKT（T308）/AKT和p-mTOR/mTOR比值均增多/升高，凋亡率和Cleaved caspase 3、Bax、E-cadherin表达均降低，差异均有统计学意义（均P<0.05）。结论 松果菊苷可抑制卵巢癌细胞的增殖、迁移、侵袭和血管生成，其机制可能与抑制PI3K/AKT/mTOR信号通路有关。

• Objective To investigate the effect of echinoside on the proliferation, migration, invasion and angiogenesis of ovarian cancer cells and its possible mechanism. Methods Ovarian serous cancer cell line SKOV3 was used as the research subjects. Cell counting kit-8 was used to detect the effect of different concentrations of echinoside (0, 6.25, 12.5, 25, 50, 100, 200, 400 μmol/L) on cell viability. SKOV3 cells were randomly divided into control group, low-dose echinacoside group (25 μmol/L), middle-dose echinacoside group (50 μmol/L), high-dose echinacoside group (100 μmol/L) and echinacoside high-dose+740Y-P(PI3K agonist, 100 μmol/L) group. Colony formation assay, TUNEL staining and Transwell assay were used to detect cell proliferation, apoptosis, migration and invasion. Angiogenesis was detected by tube formation assay. Western blotting was used to detect the expressions of proliferation, apoptosis, invasion, angiogenesis and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway related proteins. Results The results of cell counting kit-8 showed that with the increase of echinoside concentration, the survival rate of SKOV3 cells decreased gradually, which were （100.0±5.5）%,（98.3±3.3）%,（93.3±3.4）%,（58.3±3.8）%,（42.4±4.6）%,（28.8±2.2）%,（19.4±3.4）%,（6.4±1.5）%, respectively, in a concentration dependent manner (P<0.05). Compared with the control group, the number of SKOV3 cell colonies, migration, and invasive cells, the relative circular tubular structure formation ratio, the expression of cell proliferation antigen Ki-67, B-cell lymphoma 2(Bcl-2), N-cadherin, and vascular endothelial growth factor A(VEGFA), the phosphorylated(p)-PI3K/PI3K, p-AKT(S473)/AKT, p-AKT(T308)/AKT, and p-mTOR/mTOR ratios all gradually decreased in the low, medium, and high-dose echinacoside groups, while the apoptosis rate and expression of activate cysteine protease 3(Cleaved caspase 3), Bcl-2 related X protein(Bax), and E-cadherin gradually increased, showing a concentration-dependent effect, with statistically significant differences (all P<0.05). Compared with the high-dose echinacoside group, the number of SKOV3 cell colonies, migration, and invasive cells, the relative circular tubular structure formation ratio, the expression of Ki-67, Bcl-2, N-cadherin, and VEGFA, the p-PI3K/PI3K, p-AKT(S473)/AKT, p-AKT(T308)/AKT, and p-mTOR/mTOR ratios all increased/rised in the echinacoside high-dose+740Y-P group, while the apoptosis rate and expression of Cleaved caspase 3, Bax, and E-cadherin all decreased, showing a statistically significant difference(all P<0.05). Conclusion Echinoside can inhibit the proliferation, migration, invasion and angiogenesis of ovarian cancer cells, and its mechanism may be related to the inhibition of PI3K/AKT/mTOR signaling pathway.