许媛媛, 卢伊凝, 马士杰. 白头翁皂苷对胃癌细胞HGC-27增殖、迁移和侵袭的影响及可能机制[J]. 实用临床医药杂志, 2023, 27(9): 32-38. DOI: 10.7619/jcmp.20223345
引用本文: 许媛媛, 卢伊凝, 马士杰. 白头翁皂苷对胃癌细胞HGC-27增殖、迁移和侵袭的影响及可能机制[J]. 实用临床医药杂志, 2023, 27(9): 32-38. DOI: 10.7619/jcmp.20223345
XU Yuanyuan, LU Yining, MA Shijie. Influence of pulsatilla saponin on proliferation, migrationand invasion of gastric cancer HGC-27 cells and its possible mechanism[J]. Journal of Clinical Medicine in Practice, 2023, 27(9): 32-38. DOI: 10.7619/jcmp.20223345
Citation: XU Yuanyuan, LU Yining, MA Shijie. Influence of pulsatilla saponin on proliferation, migrationand invasion of gastric cancer HGC-27 cells and its possible mechanism[J]. Journal of Clinical Medicine in Practice, 2023, 27(9): 32-38. DOI: 10.7619/jcmp.20223345

白头翁皂苷对胃癌细胞HGC-27增殖、迁移和侵袭的影响及可能机制

Influence of pulsatilla saponin on proliferation, migrationand invasion of gastric cancer HGC-27 cells and its possible mechanism

  • 摘要:
    目的  探讨白头翁皂苷对胃癌细胞HGC-27增殖、迁移和侵袭的影响及其可能机制。
    方法  采用不同浓度白头翁皂苷(12.5、25.0、50.0 μmol/L)处理人胃癌细胞HGC-27, 分别设为低白头翁皂苷组、中白头翁皂苷组、高白头翁皂苷组,另将正常培养的HGC-27细胞设为对照组。将si-NC、si-circNRIP1、pcDNA、pcDNA-circNRIP1转染至HGC-27细胞,分别设为si-NC组、si-circNRIP1组、pcDNA组、pcDNA-circNRIP1组。向HGC-27细胞中转染pcDNA、pcDNA-circNRIP1, 均加入50.0 μmol/L白头翁皂苷培养,分别记为白头翁皂苷+pcDNA组、白头翁皂苷+pcDNA-circNRIP1组。通过CCK-8法、平板克隆形成实验和Transwell实验分别检测细胞增殖、克隆形成和迁移、侵袭能力; 采用实时荧光定量聚合酶链反应(qRT-PCR)法检测环状RNA核受体相互作用蛋白1 (circNRIP1)表达量; 采用蛋白质印迹法(Western blot)检测E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)的蛋白表达情况。
    结果  对照组、低白头翁皂苷组、中白头翁皂苷组、高白头翁皂苷组细胞增殖抑制率、E-cadherin蛋白水平依次升高,细胞克隆形成数量、迁移数量、侵袭数量依次减少, N-cadherin蛋白水平、circNRIP1表达量依次降低,差异均有统计学意义(P < 0.05); pcDNA-circNRIP1组circNRIP1表达量高于pcDNA组, si-circNRIP1组circNRIP1表达量低于si-NC组,差异有统计学意义(P < 0.05); si-circNRIP1组细胞增殖抑制率和E-cadherin蛋白水平高于si-NC组, N-cadherin蛋白水平低于si-NC组,细胞克隆形成数量、迁移数量和侵袭数量少于si-NC组,差异均有统计学意义(P < 0.05); 白头翁皂苷+pcDNA-circNRIP1组细胞增殖抑制率和E-cadherin蛋白水平低于白头翁皂苷+pcDNA组, N-cadherin蛋白水平高于白头翁皂苷+pcDNA组,细胞克隆形成数量、迁移数量和侵袭数量多于白头翁皂苷+pcDNA组,差异有统计学意义(P < 0.05)。
    结论  白头翁皂苷可抑制胃癌细胞增殖、迁移、侵袭活性且呈现浓度依赖性,其作用机制或与下调circNRIP1表达有关。

     

    Abstract:
    Objective  To explore the influence of pulsatilla saponin on proliferation, migration and invasion of gastric cancer HGC-27 cells and its possible mechanism.
    Methods  Human gastric cancer HGC-27 cells were treated with different doses of pulsatilla saponin (12.5, 25.0, 50.0 μmol/L) and named as low-dose pulsatilla saponin group, medium-dose pulsatilla saponin group and high-dose pulsatilla saponin group, and the normal cultured HGC-27 cells were named as control group. The si-NC, si-circNRIP1, pcDNA and pcDNA-circNRIP1 were transfected into HGC-27 cells, and were named as si-NC group, si-circNRIP1 group, pcDNA group and pcDNA-circNRIP1 group respectively. HGC-27 cells were transfected with pcDNA and pcDNA-circNRIP1 and cultured with 50.0 μmol/L pulsatilla saponin, and were named as pulsatilla saponin plus pcDNA group and pulsatilla saponin plus pcDNA-circNRIP1 group respectively. CCK-8 assay, colony formation assay and Transwell assay were used to detect the proliferation, clone formation and migration, and invasion respectively; quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of circular RNA nuclear receptor interacting protein 1 (circNRIP1); Western blot was used to detect the expressions of E-cadherin and N-cadherin proteins.
    Results  In the control group, low-dose pulsatilla saponin group, medium-dose pulsatilla saponin group and high-dose pulsatilla saponin group, the inhibition rates of cell proliferation and the levels of E-cadherin protein significantly increased gradually, the number of cell clone formation, migration and invasion significantly decreased gradually, and the expressions of N-cadherin protein and circNRIP1 significantly decreased gradually (P < 0.05); the expression level of circNRIP1 in the pcDNA-circNRIP1 group was significantly higher than that in the pcDNA group, while the expression level of circNRIP1 in the si-circNRIP1 group was significantly lower than thatin the si-NC group (P < 0.05); the inhibition rate of cell proliferation and level of E-cadherin protein in the si-circNRIP1 group were significantly higher than those in the si-NC group, while the level of N-cadherin protein and the number of cell clone formation, migration and invasion were significantly lower than those in the si-NC group (P < 0.05); the inhibition rate of cell proliferation and level of E-cadherin protein in the pulsatilla saponin plus pcDNA-circNRIP1 group were significantly lower than those in the pulsatilla saponin plus pcDNA group, while the level of N-cadherin protein and the number of cell clone formation, migration and invasion were significantly higher than those in the pulsatilla saponin plus pcDNA group (P < 0.05).
    Conclusion  Pulsatilla saponin can inhibit the proliferation, migration and invasion of gastric cancer cells in a dose-dependent manner, and its mechanism may be associated with down-regulation of circNRIP1 expression.

     

/

返回文章
返回