黄芪甲苷通过调控TGF-β/Smad信号通路抑制胃癌的侵袭、迁移及上皮间质转化的研究OACSTPCD
Study on Astragaloside Inhibiting the Invasion,Migration and Epithelial-Mesenchymal Transformation of Gastric Cancer by Regulating TGF-β/Smad Signaling Pathway
目的:探讨黄芪甲苷通过TGF-β/Smad信号通路对胃癌侵袭、迁移及上皮间质转化(EMT)相关分子表达的影响.方法:通过划痕和Transwell实验检测黄芪甲苷对胃癌细胞侵袭和迁移的影响.采用qRT-PCR和Western-blot技术分别检测黄芪甲苷对TGF-β/Smad信号通路中关键基因及EMT相关标志物mRNA和蛋白表达水平的影响.结果:划痕实验结果发现,低浓度的黄芪甲苷干预后的两株胃癌细胞愈合率明显低于对照组,差异有统计学意义(P<0.05,P<0.01),并且随着黄芪甲苷的浓度增加,愈合程度逐渐减弱.Transwell迁移和侵袭实验结果发现,黄芪甲苷干预两株胃癌细胞后穿过小室的细胞数明显低于对照组,差异有统计学意义(P<0.05,P<0.01).qRT-PCR和Western-blot实验结果发现,黄芪甲苷组的TGF-β1、TGF-β2、TGF-βR2、p-Smad2、p-Smad3 和Smad4 蛋白表达水平明显低于对照组(P<0.05,P<0.01),黄芪甲苷组和对照组之间 Smad2、Smad3 的蛋白表达水平以及TGF-β1、TGF-β2、TGF-βR2、Smad2、Smad3 和Smad4 的mRNA水平差异无统计学意义(P>0.05).黄芪甲苷组α-平滑肌肌动蛋白(α-SMA)、I型胶原(Collagen I)、N-钙黏着蛋白(N-cadherin)、细胞角蛋白18(Cytokeratin18)、Claudin7 蛋白(Claudin7)、闭锁小带蛋白-1(ZO-1)、波形蛋白(Vimentin)、锌指转录因子 1(Snail1)、锌指转录因子 2(Snail2)、纤连蛋白 1(FN1)、Twist 相关蛋白 2(Twist2)的mRNA和蛋白表达水平低于对照组(P<0.05,P<0.01),E-钙黏着蛋白(E-cadherin)的mRNA和蛋白表达水平高于对照组,差异有统计学意义(P<0.05,P<0.01).结论:黄芪甲苷通过TGF-β/Smad信号通路调控胃癌细胞的EMT,从而抑制胃癌细胞的侵袭和迁移.
Objective:To investigate the effects of astragaloside on the invasion,migration and the expression of epithelial mesenchymal transition(EMT)related molecules of gastric cancer through TGF-β/Smad signaling pathway.Methods:The effects of astragaloside on invasion and migration of gastric cancer cells were detected by scratch and transwell assay.The effects of astragaloside on mRNA and protein expression levels of key genes and EMT-related markers in TGF-β/Smad signaling pathway were detected through qRT-PCR and Western-blot,respectively.Results:The scratch test results showed that the healing rate of the two gastric cancer cells after the intervention of low concentration of astragaloside was significantly lower than that of the control group,the difference was statistically significant(P<0.05,P<0.01),and the healing degree gradually weakened with the increase of astragaloside concentration.The results of transwell migration and invasion experiment showed that the number of cells passing through the compartment after the intervention of astragaloside was significantly lower than that of the control group,and the difference was statistically significant(P<0.05,P<0.01).It was showed in qRT-PCR and Western-blot tests that the protein expression levels of TGF-β1,TGF-β2,TGF-βR2,P-SMad2,P-SMad3 and Smad4 in astragaloside group were significantly lower than those in control group(P<0.05,P<0.01).There were no significant differences in protein expression levels of Smad2 and Smad3 and mRNA levels of TGF-β1,TGF-β2,TGF-βR2,Smad2,Smad3 and Smad4 between astragaloside group and control group(P>0.05).The levels of a-SMA,Collagen 1,N-cadherin,Cytokinetin18,Claudin7,ZO-1,Vimentin,Snail1,Snail2,FN1,and mRNA,and expression levels of Twist2 mRNA and protein in astragaloside treatment group were lower than those in the control group(P<0.05,P<0.01),while the mRNA and protein levels of E-cadherin were higher than those in control group,and the difference was statistically significant(P<0.05,P<0.01).Conclusion:Astragaloside regulates EMT of gastric cancer cells through TGF-β/Smad signaling pathway,thereby inhibiting the invasion and migration of gastric cancer cells.
陈凤琴;宁月;邵利华;王蒙;李海龙
甘肃中医药大学附属医院,甘肃 兰州 730030||甘肃中医药大学第一临床医学院,甘肃 兰州 730030甘肃中医药大学第一临床医学院,甘肃 兰州 730030||甘肃中医方药挖掘与创新转化重点实验室,甘肃 兰州 730030甘肃中医药大学附属医院,甘肃 兰州 730030||甘肃中医药大学第一临床医学院,甘肃 兰州 730030||甘肃中医方药挖掘与创新转化重点实验室,甘肃 兰州 730030
中医学
黄芪甲苷TGF-β/Smad胃癌EMT侵袭迁移
AstragalosideTGF-β/SmadGastric cancerEMTInvasionMigration
《中医药学报》 2024 (010)
16-24 / 9
甘肃省自然科学基金项目(20JR5RA189);甘肃省科技厅联合科研基金一般项目(23JRRA1527);甘肃中医药大学附属医院院内创新基金项目(gzfy-2022-12)
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