威灵仙总皂苷通过miR-410-3p介导调控胶质瘤细胞的生物学活性

The total saponins of Clematidis radix et rhizoma regulate the biological activity of glioma cells through miR-410-3p mediation

目的探讨威灵仙总皂苷通过调控微小RNA(miRNA)-410-3p对胶质瘤细胞生物学活性的影响以及对第10号染色体缺失的磷酸酶和张力蛋白同源物基因/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(phosphatase and tensin homolog deleted on chromosome ten/protein kinase B/mammalian target of rapamycin,PTEN/Akt/mTOR)信号通路的调节作用。方法用不同浓度威灵仙总皂苷处理U87细胞后,用CCK-8法检测细胞存活率。将对数生长期U87细胞随机分为对照组、NC组(转染miR-410-3p NC)、inhibitor组(转染miR-410-3p inhibitor)以及inhibitor联合威灵仙总皂苷组(转染miR-410-3p inhibitor 24 h后,40μmol·L^(-1)威灵仙总皂苷处理),依次检测miR-410-3p表达水平、细胞存活率、细胞迁移和侵袭力,miR-410-3p与PTEN的靶向关系和PTEN、p-Akt、Akt、p-mTOR以及mTOR蛋白的相对表达量。结果细胞存活率随威灵仙总皂苷浓度的升高而降低(P<0.05);与NC组比较,inhibitor组划痕愈合率、侵袭细胞数量、p-Akt和p-mTOR蛋白的相对表达量升高,miR-410-3p表达水平以及PTEN蛋白的相对表达量降低(P<0.05);与inhibitor组比较,inhibitor联合威灵仙总皂苷组细胞存活率、划痕愈合率、侵袭细胞数量、p-Akt和p-mTOR蛋白相对表达量降低,miR-410-3p表达水平以及PTEN蛋白相对表达量升高(P<0.05)。结论威灵仙总皂苷可抑制胶质瘤细胞的恶性生物学行为,其可能是通过上调miR-410-3p表达,进而调控PTEN/Akt/mTOR信号通路发挥作用。

Objective To investigate the effects of the total saponins of Clematidis radix et rhizoma(TS)on the biological activity of glioma cells by regulating microRNA(miRNA)-410-3p,and on the regulation of phosphatase and tensin homolog gene/protein kinase B/mammalian target of rapamycin(PTEN/Akt/mTOR)signaling pathway with deletion of chromosome 10.Methods U87 cells were treated with different concentrations of TS,and the cell survival rate was detected by CCK-8 method.U87 cells at logarithmic growth stage were randomly divided into control group,NC group(transfected with miR-410-3p NC),inhibitor group(transfected with miR-410-3p inhibitor),and inhibitor combined with TS group(transfected with miR-410-3p inhibitor 24 hours,then treated with 40μmol·L^(-1)of TS).The expression level of miR-410-3p,cell survival rate,cell migration and invasiveness,the targeting relationship between miR-410-3p and PTEN,as well as the relative expression levels of PTEN,p-Akt,Akt,p-mTOR and mTOR protein were detected in sequence.Results The cell survival rate was decreased with the increase of TS concentration(P<0.05).Compared with NC group,the scratch healing rate,the number of invaded cells,the relative expression levels of p-Akt and p-mTOR proteins in the inhibitor group were increased,while the expression levels of miR-410-3p and the relative expression levels of PTEN protein were decreased(P<0.05).Compared with inhibitor group,the cell survival rate,scratch healing rate,number of invaded cells,relative expression levels of p-Akt and p-mTOR proteins were decreased,and the miR-410-3p expression level and relative expression level of PTEN protein were increased in inhibitor combined with TS group(P<0.05).Conclusion TS can inhibit the malignant bio-behavior of glioma cells,possibly by regulating the PTEN/Akt/mTOR signaling pathway by up-regulating the expression of miR-410-3p.