河北医学2023，Vol.29Issue(12)：1985-1992,8.DOI:10.3969/j.issn.1006-6233.2023.12.010
miR-21靶向PTEN调控AKT/FoxO1信号通路对胃癌细胞凋亡的机制研究
Mechanism of miR-21 Targeting PTEN to Regulate AKT/FoxO1 Signaling Pathway on Apoptosis in Gastric Cancer Cells
摘要
Abstract
Objective:To investigate the effect of miR-21 targeting PTEN to regulate AKT/FoxO1 sig-naling pathway on apoptosis in gastric cancer cells.Methods:Gastric cancer SGC-7901 cells were divided into miR-NC inhibitors group(SGC-7901 cells transfected with miR-NC inhibitors plasmid),miR-21 inhib-itors group(SGC-7901 cells transfected with miR-21 inhibitors plasmid),miR-21 inhibitors+sh-PTEN group(SGC-7901 cells transfected with miR-21 inhibitors and sh-PTEN plasmids);and control group(SGC-7901 cells not transfected with any plasmids).qRT-PCR was performed to detect miR-21 mRNA ex-pression in cells;CCK-8 was used to detect cell proliferation ability;Transwell assay for cell invasion shift a-bility;flow cytometry for apoptosis rate;protein blotting for PTEN,AKT,p-AKT,PI3K,p-PI3K,FoxO1 protein expression in cells;dual luciferase reporter for targeting relationship between miR-21 and PTEN.Re-sults:Compared with normal human gastric mucosal epithelial cell GES-1(1.00±0.10),the expression of miR-21(1.89±0.17)in gastric cancer cell SGC-7901 was significantly increased(P<0.05).Compared with the control group,the expression of miR-21(0.83±0.10),proliferation rate(45.31±4.92)%,number of invasions(62.34±5.83),and p-PI3K/PI3K(0.42±0.05),p-AKT/AKT(0.51±0.05)ratios were significantly reduced in the miR-21 inhibitors group,while the apoptosis rate(23.48±3.51)%,PTEN(0.98±0.10),and FoxO1(0.76±0.08)protein expression were significantly increased(P<0.001).Compared with the pcDNA-NC group,the cell proliferation rate(48.26±5.01),number of invasive cells(65.37±6.02),and p-PI3K/PI3K(0.46±0.05),p-AKT/AKT(0.55±0.06)ratios in the pcD-NA-PTEN group were significantly reduced,while the cell apoptosis rate(25.61±3.27)%and the expres-sion of PTEN(0.91±0.09)and FoxO1(0.70±0.08)proteins in the cells were increased(P<0.05).It was predicted that the 3'UTR end of PTEN had complementary binding sites with miR-21.Compared with the miR-NC group,the luciferase activity of the miR-21 group(0.32±0.03)was significantly reduced(P<0.05)when transfected with wild-type PTEN(PTEN-WT).Compared with the miR-21 inhibitors group,the miR-21 inhibitors+sh-PTEN group showed a significant increase in cell proliferation rate(90.25±9.14)%,invasion cell count(125.69±10.31),and p-PI3K/PI3K(0.80±0.08),p-AKT/AKT(0.76±0.08)ratios.The apoptosis rate(6.24±1.32)and the expression of PTEN(0.30±0.04)and FoxO1(0.38±0.05)proteins in the cells were also decreased(P<0.001).Conclusion:Knockdown of miR-21 can target negative regulation of PTEN expression,modulate AKT/FoxO1 signaling pathway,inhibit proliferation and in-vasion of gastric cancer cells,and promote apoptosis.关键词
胃癌/miR-21/PTEN/AKT/FoxO1信号通路/凋亡Key words
Gastric cancer/miR-21/PTEN/AKT/FoxO1 signaling pathway/Apoptosis引用本文复制引用
王艳花,聂亚楠,齐俊娟,季艳霞..miR-21靶向PTEN调控AKT/FoxO1信号通路对胃癌细胞凋亡的机制研究[J].河北医学,2023,29(12):1985-1992,8.基金项目
2020年度河北医学科研课题计划,(编号:20200195) （编号:20200195）
