竹节参皂苷IVa调控DNMT1表达影响肝细胞癌细胞株增殖、迁移、侵袭及干性特征研究OA
The Research of Chikusetsusaponin IVa Regulated DNMT1 Expression to Affect Proliferation,Migration,Invasion,and Stemness of Hepatocellular Carcinoma Cells
目的 探究竹节参皂苷IVa(CsIVa)调控肝细胞癌细胞生长、转移及干细胞特性的功能及机制.方法 qRT-PCR和Western blot检测DNA甲基转移酶1(DNMT1)mRNA和蛋白在肝细胞癌细胞株HepG2、Huh7、Hep3B、MHCC97-H和人肝细胞LO2中的表达.HepG2细胞分为CsIVa 0、50、100、200 μmol/L组和CsIVa 200 μmol/L+DNMT1组.CCK-8、流式细胞术和细胞成球实验检测各组细胞增殖、凋亡和干性,Transwell法检测细胞迁移和侵袭能力.Western blot检测各组细胞干性标志物(Nanog、Sox2、Oct4、KLF4和CD133)蛋白表达.结果 MHCC97-H、Hep3B、Huh7和HepG2中DNMT1 mRNA和蛋白表达水平均显著高于其在LO2中的DNMT1 mRNA和蛋白表达(tmRNA=8.10～45.09,t蛋白=4.63～22.66,均P<0.05).与CsIVa 0 μmol/L组比较,CsIVa 50、100和200 μmol/L组HepG2细胞DNMT1表达、细胞增殖、迁移、侵袭、细胞成球数显著降低,差异具有统计学意义(t=4.32～36.18,均P<0.05),Nanog、Sox2、Oct4、KLF4和CD133蛋白表达显著降低,差异具有统计学意义(t=4.09～39.27,均P<0.05),细胞凋亡率显著增加(t=7.12～18.34,均P<0.05).与CsIVa 200 μmol/L组比较,CsIVa 200 μmol/L+DNMT1组HepG2细胞DNMT1表达、细胞增殖、迁移、侵袭、细胞成球数显著增加,差异具有统计学意义(t=7.42～16.35,均P<0.05),Nanog、Sox2、Oct4、KLF4和CD133蛋白表达显著增加,差异具有统计学意义(t=10.79～24.80,均P<0.05),细胞凋亡显著降低(t=4.48,P<0.05).结论 CsIVa下调DNMT1表达而抑制肝细胞癌细胞生长和转移能力以及干细胞特性.
Objective To explore the function and mechanism of chikusetsusaponin IVa(CsIVa)in regulating the growth,metas-tasis,and stemness characteristics of hepatocellular carcinoma cells.Methods qRT-PCR and Western blot were used to detect the mRNA and protein expression of DNA Methyltransferase 1(DNMT1)in hepatocellular carcinoma cell lines HepG2,Huh7,Hep3B,and MHCC97-H,as well as in human hepatocytes LO2.HepG2 cells were divided into groups treated with 0,50,100,or 200 μmol/L CsIVa,and a group treated with 200 μmol/L CsIVa+DNMT1 overexpression.CCK-8 assay,flow cytometry,and cell spheroid formation assay were used to assess cell proliferation,apoptosis,and stemness characteristics,respectively.Transwell assay was used to evaluate cell migration and invasion abilities.Western blot was used to detect the protein expression of stemness markers(Nanog,Sox2,Oct4,KLF4,and CD133)in each group of cells.Results The expression levels of DNMT1 mRNA and protein in MHCC97-H,Hep3B,Huh7,and HepG2 were significantly higher than those in LO2(tmRNA=8.10～45.09,tprotein=4.63～22.66,all P<0.05).Compared with the CsIVa 0 μmol/L group,the CsIVa 50,100,and 200 μmol/L groups showed significantly reduced DNMT1 expression,cell proliferation,migration,invasion,and cell spheroid formation in HepG2 cells(t=4.32～36.18,all P<0.05),the protein levels of Nanog,Sox2,Oct4,KLF4,and CD133 were also significantly decreased(t=4.09～39.27,all P<0.05),and the apoptosis rate was significantly increased(t=7.12～18.34,all P<0.05).Compared with the CsIVa 200 μmol/L group,the CsIVa 200 μmol/L+DNMT1 group showed significantly increased DNMT1 expression,cell proliferation,migration,invasion,and cell spheroid formation in HepG2 cells(t=7.42～16.35,all P<0.05),the protein levels of Nanog,Sox2,Oct4,KLF4,and CD133 were also significantly increased(t=10.79～24.80,all P<0.05),and the apoptosis rate was significantly reduced(t=4.48,P<0.05).Conclusions CsIVa downregulates DNMT1 expression to inhibit the growth,and meta-static potential,and stemness characteristics of hepatocellular carcinoma cells.
冯变莹;陈静奇;郝湘文;宗鸣;陈皓;满秋红
同济大学附属上海市第四人民医院检验科,上海 200434同济大学附属上海市第四人民医院检验科,上海 200434同济大学附属上海市第四人民医院检验科,上海 200434同济大学附属上海市第四人民医院检验科,上海 200434同济大学附属上海市第四人民医院检验科,上海 200434同济大学附属上海市第四人民医院检验科,上海 200434
医药卫生
竹节参皂苷IVa肝细胞癌DNA甲基转移酶1增殖迁移侵袭干性特征
Chikusetsusaponin IVahepatocellular carcinomaDNA methyltransferase 1proliferationmigrationinvasionstemness characteristics
《现代检验医学杂志》 2026 (5)
46-51,6
上海市第四人民医院科研启动专项(sykyqd02201).
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