南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (8): 1315-1321.doi: 10.12122/j.issn.1673-4254.2023.08.07

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桂枝茯苓胶囊通过调控NF-κB通路抑制卵巢癌细胞的迁移和诱导卵巢癌细胞的凋亡

郭晓娟，陈丽平，吕 芹，杜瑞娟，罗 琴，张 阳，卞 华，韩 立   

  1. 南阳理工学院张仲景国医学院，河南省张仲景方药与免疫调节重点实验室，河南 南阳 473004；南阳医学高等专科学校中医系，河南 南阳 473061
  • 出版日期:2023-08-20 发布日期:2023-09-13

Guizhi Fuling Capsule inhibits migration and induces apoptosis of human ovarian cancer cells by regulating the NF-κB signaling pathway

GUO Xiaojuan, CHEN Liping, LÜ Qin, DU Ruijuan, LUO Qin, ZHANG Yang, BIAN Hua, HAN Li   

  1. Zhang Zhongjing School of Chinese Medicine, Henan Key Laboratory of ZHANG Zhongjing Formulae and Herbs for Immunoregulation, Nanyang Institute of Technology, Nanyang 473004, China; Department of Chinese Medicine, Nanyang Medical College, Nanyang 473061, China
  • Online:2023-08-20 Published:2023-09-13

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摘要： 目的 研究桂枝茯苓胶囊（GFC）抑制人卵巢癌转移的作用和潜在机制。方法 60只Wistar大鼠随机分为4组，15只/组：给药组分别灌胃给予4、8、16 g·kg-1·d-1剂量的GFC悬液5 d，分别制备GFC低、中、高剂量含药血清，空白组给予相同体积的生理盐水制备空白血清；卵巢癌耐顺铂SKOV3/DDP细胞分为空白血清组、不同剂量GFC含药血清组和NF-κB抑制剂SN50阳性对照组。采用划痕实验检测含药血清对迁移的影响，采用流式细胞术检测含药血清对凋亡的影响，采用RT-qPCR和Western blot检测含药血清对细胞中上皮表型E-钙粘蛋白（CDH1）、间质表型N-钙黏蛋白（CDH2）、caspase 3和NF-κB mRNA和蛋白表达的影响。采用ATAC-seq检测染色质开放区域CDH1、CDH2、caspase 3和NF-κB 基因表达差异。结果 GFC含药血清可剂量依赖性地抑制SKOV3/DDP细胞迁移，增加细胞凋亡（P<0.05，P<0.01）。随着含药血清剂量增加，其对CDH2和NF-κB的抑制作用，及其对CDH1和caspase 3的诱导作用均逐渐增强（P<0.05，P<0.01）。高剂量含药血清抑制SKOV3/DDP细胞迁移，诱导凋亡、caspase 3 mRNA和蛋白以及CDH1 mRNA表达，抑制CDH2和NF-κB mRNA和蛋白表达的作用与SN50相当（P>0.05），但诱导CDH1蛋白表达的作用弱于SN50（P<0.01）。含药血清可减弱染色质开放区域NF-κB、CDH2基因表达，增强CDH1表达，但对caspase 3无影响。结论 GFC含药血清抑制SKOV3/DDP迁移作用可能与其诱导凋亡、caspase 3和CDH1表达，抑制CDH2和NF-κB表达，调控染色质开放区域NF-κB、CDH2和CDH1表达有关。

关键词: 卵巢癌；桂枝茯苓胶囊；核因子κB；E-钙粘蛋白；N-钙黏蛋白；上皮间质转化；半胱氨酸蛋白酶-3

Abstract: Objective To study the inhibitory effect of Guizhi Fuling Capsule (GFC) on migration of human ovarian cancer cells and explore the possible mechanism. Methods Sixty Wistar rats were randomized into 4 groups for daily gavage of saline or 4, 8, or 16 g/kg GFC suspension for 5 days to prepare blank and low-, medium- and high-dose GFC-medicated sera. Cisplatin-resistant ovarian cancer SKOV3/DDP cells were treated with these sera with nuclear factor-κB (NF-κB) inhibitor SN50 as the positive control, and the changes in migration ability and apoptosis of the cells were examined using scratch assay and flow cytometry, respectively; the changes in the mRNA and protein expressions of CDH1, CDH2, caspase 3 and NF- κB were detected using RT- qPCR and Western blotting. ATAC- seq was used to analyze the changes in expressions of CDH1, CDH2, caspase 3 and NF-κB genes in the open chromatin. Results Treatment with GFC-medicated sera dose-dependently inhibited the migration (P<0.05), increased apoptosis (P<0.01), inhibited CDH2 and NF-κB mRNA expression (P<0.05), and enhanced caspase 3 and CDH1 mRNA expressions (P<0.01) in SKOV3/DDP cells. The effects of high-dose GFC-medicated serum were comparable to those of SN50 (P>0.05), but its effect for enhancing DH1 protein expression was weaker than that of SN50 (P<0.01). GFC-medicated sera significantly lowered the expressions of NF-κB and CDH2 and increased CDH1 expression in the open chromatin without obviously affecting caspase 3 expression. Conclusion GFC- medicated sera inhibits the migration ability of SKOV3/DDP cells possibly by promoting cell apoptosis and caspase 3 and CDH1 expressions, inhibiting CDH2 and NF-κB expressions, and regulating the expressions of NF-κB, CDH2 and CDH1 in the open chromatin.

Key words: ovarian cancer; Guizhi Fuling Capsule; nuclear factor- kappa B; E-cadherin; N-cadherin; epithelial-mesenchymal transition; caspase 3