扁蒴藤素调节Shh/Gli1信号通路对宫颈癌HeLa细胞增殖、凋亡和血管生成拟态的影响OA北大核心CSTPCD
Effects of pristimerin on the proliferation,apoptosis and vasculogenic mimicry of cervical cancer HeLa cells by regulating the Shh/Gli1 signaling pathway
目的:探讨扁蒴藤素(Pris)调节Shh/Gli1信号通路对宫颈癌HeLa细胞增殖、凋亡和血管生成拟态(VM)的影响及其机制.方法:采用MTT法检测不同浓度Pris对宫颈癌HeLa细胞增殖的抑制作用,以选取合适的干预浓度.将HeLa细胞分为对照组、环巴胺组、Pris组、Pris+pc-NC组和Pris+pc-Shh组.采用MTT法、EdU法检测各组细胞的增殖能力,Transwell小室法、流式细胞术检测各组细胞的迁移及侵袭能力和细胞凋亡率,体外血管生成实验观察VM形成情况,qPCR法检测各组细胞中Shh和Gli1 mRNA表达水平,WB法检测细胞中血管内皮生长因子A(VEGF-A)、血管内皮钙黏素(VE-cadherin)、Ki-67、caspase-3及与Shh/Gli1信号通路相关蛋白表达水平.结果:0.25～2.5 μmol/L的Pris对HeLa细胞增殖均有显著抑制作用,选择1.5 μmol/L的Pris进行后续实验.对照组细胞形成良好的管腔结构,与对照组相比,环巴胺组、Pris组和Pris+pc-NC组HeLa细胞管腔结构被明显破坏,细胞增殖活力和增殖率、迁移及侵袭细胞数目、Shh和Gli1 mRNA、VEGF-A、VE-cadherin、Ki-67、Shh、Gli1蛋白表达均显著降低(均P<0.05),细胞凋亡率和caspase-3表达均显著升高(均P<0.05);环巴胺组与Pris组HeLa细胞各项检测指标比较差异均无统计学意义(均P>0.05);与Pris+pc-NC组相比,Pris+pc-Shh组细胞管腔结构形成明显改善,细胞增殖活力和增殖率、迁移及侵袭细胞数、Shh和Gli1 mRNA、VEGF-A、VE-cadherin、Ki-67、Shh、Gli1蛋白表达均显著升高(均P<0.05),细胞凋亡率和caspase-3表达均显著降低(均P<0.05).结论:Pris抑制宫颈癌HeLa细胞的增殖、迁移与侵袭和VM的形成并促进细胞凋亡,可能与阻断Shh/Gli1信号通路有关.
Objective:To investigate the effects of pristimerin(Pris)on the proliferation,apoptosis and vasculogenic mimicry(VM)of cervical cancer HeLa cells by regulating the Shh/Gli1 signaling pathway.Methods:MTT method was used to detect the inhibitory effects of Pris in different concentrations on the proliferation of cervical-cancer HeLa cells to select appropriate intervention concentration.Cervical cancer HeLa cells were grouped into the control group,the cyclopamine group,the Pris group,the Pris+pc-NC group and the Pris+pc-Shh group.MTT and EdU were applied to detect the proliferation abilities of cells in each group.Transwell chamber method and flow cytometry were used to detect cell migration and invasion abilities and cell apoptosis rate.In vitro angiogenesis experiments were used to observe the formation of VM.qPCR was used to detect Shh and Gli1 mRNA expression levels in each group.Western blotting was used to detect the expression levels of vascular endothelial growth factor A(VEGF-A),vascular endothelial cadherin(VE-cadherin),Ki-67,caspase-3,and proteins related to the Shh/Gli1 signaling pathway.Results:0.25-2.5 mol/L Pris significantly inhibited the proliferation of HeLa cells,and 1.5 mol/L was selected for subsequent experiments.The cells in the control group formed a good lumen structure.Compared with the control group,the lumen structures of HeLa cells in the cyclopamine group,the Pris group and the Pris+pc-NC group were significantly damaged.Cell proliferation activity,proliferation rate,numbers of migration and invasion,expression levels of Shh and Gli1 mRNA,and expression levels of VEGF-A,VE-cadherin,Ki-67,Shh and Gli1 proteins were significantly decreased(all P<0.05).The apoptosis rate and the expression level of caspase-3 were significantly increased(all P<0.05).There was no significant difference in HeLa cell detection indicators between the cyclopamine group and the Pris group(all P>0.05).Compared with the Pris+pc-NC group,the formation of cell lumen structure in the Pris+pc-Shh group was significantly improved.The cell proliferation activity and proliferation rate,the numbers of migration and invasion cells,the expressions of Shh and Gli1 mRNA,and the expressions of VEGF-A,VE-cadherin,Ki-67,Shh and Gli1 proteins were significantly increased(all P<0.05).The apoptosis rate and the expression level of caspase-3 were significantly decreased(all P<0.05).Conclusion:Pris can inhibit the proliferation,migration,invasion and VM formation of cervical cancer HeLa cells,and promote cell apoptosis,which may be related to blocking the Shh/Gli1 signaling pathway.
罗健玮;黄泓轲;胡艳丽
乐山职业技术学院 中医康养学院,四川 乐山 614000乐山职业技术学院 生物医药学院,四川 乐山 614000重庆大学附属黔江医院 妇科,重庆 400030
临床医学
扁蒴藤素宫颈癌HeLa细胞Shh/Gli1信号通路血管生成拟态
pristimerin(Pris)cervical cancerHeLa cellShh/Gli1 signaling pathwayvasculogenic mimicry(VM)
《中国肿瘤生物治疗杂志》 2024 (007)
687-693 / 7
乐山市科技计划项目(No.20SZD004)
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