三七总皂苷抑制胃癌进程的体外研究

Inhibition of total saponins in panax notoginseng on progression of gastric cancer in vitro

目的:探究三七总皂苷对人胃癌细胞系SGC-790、BGC-823及MKN-4的增殖、迁移、侵袭和凋亡等肿瘤生物学行为的影响及可能涉及的机制。方法:通过CCK8方法研究三七皂总苷对胃癌细胞系增殖能力的影响,通过Transwell侵袭实验研究三七总皂苷对胃癌细胞系侵袭能力的影响,通过划痕实验研究三七总皂苷对胃癌细胞系迁移能力的影响,通过流式方法检测三七总皂苷对胃癌细胞系凋亡水平的影响;采用Western blot检测三七总皂苷对胃癌细胞系中Vemintin、SNAIL、MMP2、MMP9、P21、Caspase-3、BAX和β-catenin等蛋白表达水平的影响;采用免疫荧光方法检测三七总皂苷对胃癌细胞系SGC-790中β-catenin的细胞空间分布的影响。结果:CCK8实验结果证实三七总皂苷可以显著抑制胃癌细胞系的增殖能力(P<0.05),Transwell侵袭实验证实三七总皂苷可以抑制胃癌细胞系的侵袭能力(P<0.05),划痕实验证明三七总皂苷可以抑制胃癌细胞系的迁移能力(P<0.05),凋亡实验证实三七总皂苷可以诱导胃癌细胞系凋亡水平增高(P<0.05);Western blot方法证实三七总皂苷可以下调Vemintin、SNAIL、MMP2和MMP9的蛋白表达量,并上调P21、Caspase-3和BAX的蛋白水平,可以下调细胞核内β-catenin的蛋白水平(P<0.05);免疫荧光结果证实,三七总皂苷可以减少β-catenin的细胞核内荧光强度,改变β-catenin的细胞内空间分布(P<0.05)。结论:三七总皂苷可以显著抑制胃癌细胞系的增殖、侵袭、迁移能力,并诱导胃癌细胞系发生凋亡。其机制是三七总皂苷在胃癌细胞内可以抑制WNT/β-catenin通路,其可能是一种治疗胃癌的新药物。

Objective:To investigate the effects of total saponins of panax notoginseng on the biological behaviors of human gastric cancer cell lines such as proliferation,migration,invasion and apoptosis of SGC-790,BGC-823 and MKN-4 and the possible mechanisms involved.Methods:Total notoginseng saponin glycosides were studied by CCK8 method research of gastric cancer cell line proliferation,the influence of notoginseng were detected by Transwell invasion experiment study,the effects of total saponins in gastric cancer cell line attack through experiments scratches,total saponin panax notoginseng effect on gastric cancer cell apoptosis level was detected by flow cytometry.The effect of total saponins of panax notoginseng on expression levels of Vemintin,SNAIL,MMP2,MMP9,P21,caspase-3,BAX andβ-catenin in gastric cancer cell lines was detected by Western blot.The effect of total saponins of panax notoginseng on the spatial distribution ofβ-catenin in gastric cancer cell line SGC-790 was detected by immunofluorescence.Results:CCK8 experimental results confirmed notoginseng total saponin could significantly inhibit the proliferation of gastric cancer cell line(P<0.05),Transwell invasion experiment confirmed notoginseng total saponin could inhibit the invasive ability of gastric cancer cell line(P<0.05),scratch experiments showed notoginseng total saponin could inhibit the migration ability of gastric cancer cell line(P<0.05)and apoptosis experiments confirmed notoginseng total saponin could induce apoptosis of gastric cancer cell line level increased(P<0.05).Western blot confirmed that the total saponins of panax notoginseng could down-regulate the protein expression levels of Vemintin,SNAIL,MMP2 and MMP9,up-regulate the protein levels of P21,Caspase-3 and BAX,and down-regulate the protein levels ofβ-catenin in the nucleus(P<0.05).Immunofluorescence results confirmed that total saponins of panax notoginseng could reduce the intracellular fluorescence intensity ofβ-catenin and change the intracellular spatial distribution ofβ-catenin(P<0.05).Conclusion:Total saponins of panax notoginseng can significantly inhibit the proliferation,invasion and migration of gastric cancer cell lines,and induce the apoptosis of gastric cancer cell lines.The mechanism is that total saponins of panax notoginseng can inhibit WNT/β-catenin pathway in gastric cancer cells,and total saponins of panax notoginseng may be a new drug for the treatment of gastric cancer.