天然产物研究与开发 ›› 2025, Vol. 37 ›› Issue (11): 2012-2017.doi: 10.16333/j.1001-6880.2025.11.004 cstr: 32307.14.1001-6880.2025.11.004

• 研究论文 • 上一篇    下一篇

基于Wnt/β-catenin通路探究通关藤C21甾体皂苷改善卵巢癌细胞顺铂耐药的分子机制

李观荣,刘黎蓝,彭素娟,郑石洲*   

  1. 广东医科大学附属第二医院,湛江 524003
  • 出版日期:2025-11-27 发布日期:2025-11-26
  • 基金资助:
    广东省基础与应用基础研究基金(2019A1515110386)

Study on the molecular mechanism involved in reducing cisplatin resistance of Marsdenia tenacissima C21 steroid saponin against ovarian cancer cells based on Wnt/β-catenin pathway 

LI Guan-rong,LIU Li-lan,PENG Su-juan,ZHENG Shi-zhou*   

  1. The Second Affiliated Hospital of Guangdong Medical University,Zhanjiang,524003,China 
  • Online:2025-11-27 Published:2025-11-26

摘要:

基于无翅型MMTV整合位点家族(wingless-type MMTV integration site family,Wnt)/β-连环蛋白(β-catenin)通路分析通关藤C21甾体皂苷(Marsdenia tenacissima C21 steroid saponins,MTC21)对卵巢癌细胞顺铂耐药的影响及相关分子机制。取对数期生长卵巢癌顺铂(cisplatin,DDP)耐药细胞株SKOV3/DDP细胞,采用CCK-8法检测不同浓度DDP(0、1、2、4、8、16 μg/mL)干预下SKOV3/DDP细胞的增殖情况。分别设置对照组(control group,Con)、顺铂组(DDP,2 μg/mL)及MTC21低剂量组(MTC21 in Low-dose group,L-MTC21)、MTC21中剂量组(MTC21 in Medium-dose group,M-MTC21)、MTC21高剂量组(MTC21 in High-dose group,H-MTC21)(分别给予2 μg/mL的DDP及0.05、0.2、0.8 mg/mL MTC21干预)。采用流式细胞法检测各组细胞凋亡情况,采用实时荧光定量聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)及蛋白免疫印迹(Western blot,WB)法检测Wnt、β-catenin mRNA和蛋白的表达情况。量效曲线显示,Con组DDP对SKOV3/DDP细胞的半数抑制浓度(50% concentration of inhibition,IC50)为10.9 μg/mL,而L-MTC21、M-MTC21及H-MTC21组的IC50分别为9.1、8.3、6.5 μg/mL,随MTC21浓度升高DDP对细胞的IC50值降低,其中H-MTC21组IC50值最低。与Con组比较,DDP组、L-MTC21、M-MTC21及H-MTC21组细胞凋亡率升高(P<0.05),Wnt和β-catenin mRNA及蛋白表达量降低(P<0.05);与DDP组比较,L-MTC21、M-MTC21及H-MTC21组细胞凋亡率升高(P<0.05),Wnt和β-catenin mRNA及蛋白表达量降低(P<0.05);与L-MTC21组比较,M-MTC21组、H-MTC21组细胞凋亡率升高(P<0.05),Wnt和β-catenin mRNA及蛋白表达量降低(P<0.05);与M-MTC21组比较,H-MTC21组细胞凋亡率升高(P<0.05),Wnt和β-catenin mRNA及蛋白表达量降低(P<0.05)。综上所述,MTC21可增加卵巢癌SKOV3/DDP细胞对DDP化疗的敏感性,其作用机制可能与抑制Wnt/β-catenin通路有关。

关键词: 卵巢癌, 通关藤C21甾体皂苷, 顺铂耐药, β-连环蛋白

Abstract:

Based on the wingless MMTV integration site family (Wnt)/β-catenin (β-catenin) pathway, the effect of Marsdenia tenacissima C21 steroid saponin (MTC21) on cisplatin resistance in ovarian cancer cells and related molecular mechanisms were analyzed. SKOV3/DDP cell line with cisplatin-resistant ovarian cancer was selected. The proliferation of SKOV3/DDP cells under different concentrations of DDP (0, 1, 2, 4, 8, 16 μg/mL) was detected using CCK-8. The control group (Con), cisplatin group (DDP; 2 μg/mL) and the MTC21 in Low-dose group (L-MTC21), MTC21 in Medium-dose group (M-MTC21), and MTC21 in High-dose group (H-MTC21) (2 μg/mL DDP and 0.05, 0.2, 0.8 mg/mL MTC21 interventions) were set up. Flow cytometry was used to detect the apoptosis of each group, and the mRNA and protein expressions of Wnt and β-catenin were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB). The dose-response curve showed that the 50% concentration of inhibition (IC50) of DDP against SKOV3/DDP cells in the Con group was 10.9 μg/mL. The IC50 of the L-MTC21, M-MTC21 and H-MTC21 groups were 9.1, 8.3 and 6.5 μg/mL, respectively. With the increase of the concentration of MTC21, the IC50 value of DDP decreased on cells, and the IC50 value of H-MTC21 group was the lowest. Compared with the Con group, the apoptosis rate of the DDP group, L-MTC21, M-MTC21 and H-MTC21 groups increased (P<0.05), and the mRNA and protein expressions of Wnt and β-catenin decreased (P<0.05). Compared with the DDP group, the apoptosis rate of the L-MTC21, M-M-MTC21 and H-MTC21 groups increased (P<0.05), and the mRNA and protein expressions of Wnt and β-catenin decreased (P<0.05). Compared with the L-MTC21 group, the apoptosis rate of the M-MTC21 group and H-MTC21 group increased (P<0.05), and the mRNA and protein expressions of Wnt and β-catenin decreased (P<0.05). Compared with the M-MTC21 group, the apoptosis rate of the H-MTC21 group was increased (P<0.05), and the mRNA and protein expressions of Wnt and β-catenin were decreased (P<0.05). In conclusion, MTC21 can increase the sensitivity of ovarian cancer SKOV3/DDP cells to DDP chemotherapy, and its mechanism of action may be related to the inhibition of Wnt/β-catenin pathway.

Key words: ovarian cancer, Marsdenia tenacissima C21 steroid saponin, cisplatin resistance, β-catenin

中图分类号:  R282 R937.31