环境与职业医学2025，Vol.42Issue(4)：467-474,8.DOI:10.11836/JEOM24244
亚砷酸钠通过调控ER-PI3K/AKT信号通路影响甲状腺正常细胞增殖与凋亡
Sodium arsenite influences proliferation and apoptosis in normal thyroid cells via modulation of ER-PI3K/AKT signaling pathway
摘要
Abstract
[Background]Recent advances in understanding the toxic effects of inorganic arsenic have revealed that arsenic exposure impacts multiple endocrine organs,thereby altering their functions.However,the mechanisms underlying arsenic-induced thyroid injury remain unclear. [Objective]To investigate the mechanisms by which sodium arsenite(NaAsO₂)affects the proliferation and apoptosis of normal thyroid cells(Nthy-ori3-1)through the estrogen receptor(ER)-phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway. [Methods]Nthy-ori3-1 cells were cultured in vitro and divided into the following groups:a control group(complete medium without drugs,0 μmol·L-1),and NaAsO₂-treated groups at 1,2,and 4 μmol·L-1.Additionally,1 μmol·L-1 of the ER inhibitor ICI182780 was used to intervene in the NaAsO₂ exposure groups,resulting in the following combinations:1 μmol·L-1 NaAsO₂+ICI182780,2 μmol·L-1 NaAsO₂+ICI182780,and 4 μmol·L-1 NaAsO₂+ICI182780.The median lethal concentration of NaAsO₂ was determined using cell viability assay.Cell viability was assessed at 24,36,and 48 h using Cell Counting Kit-8(CCK-8)assay.Colony formation ability was evaluated via plate cloning assay.Apoptosis was detected using Hoechst 33342 staining.Protein and mRNA expression levels of ERα,ERβ,c-MYC,Bax,Bcl-2,PI3K,p-PI3K,AKT,and p-AKT were measured using Western blot(WB)and real-time quantitative PCR(qRT-PCR),respectively. [Results]The median lethal concentration of NaAsO₂ was determined to be 4.034 μmol·L-1.CCK-8 assay at 24,36,and 48 h revealed that,compared with the control group,the 1,2,and 4 μmol·L-1 NaAsO₂ groups significantly inhibited Nthy-ori3-1 cell proliferation(P<0.001).The plate cloning assays demonstrated a concentration-dependent reduction in colony formation ability(P<0.001).Following the ICI182780 intervention,the cell viability and colony formation ability in the 1,2,and 4 μmol·L-1 NaAsO₂ groups were significantly restored compared with the corresponding NaAsO₂-only groups(P<0.001,P<0.01).The Hoechst 33342 staining indicated that compared with the control group,the nuclear staining intensity and apoptosis levels in the 1,2,and 4 μmol·L-1 NaAsO₂ groups increased in a concentra-tion-dependent manner(P<0.001).However,the ICI182780 intervention reduced the apoptosis levels in the NaAsO₂-treated groups compared with their NaAsO₂-only counterparts(P<0.001).The WB analysis showed that,compared with the control group,the protein expression of ERα,ERβ,c-MYC,Bcl-2,p-PI3K,and p-AKT in the 1,2,and 4 μmol·L-1 NaAsO₂ groups decreased manner(P<0.001,P<0.01),while the Bax expression increased in a concentration-dependent(P<0.001);the PI3K and AKT protein levels showed no significant dif-ferences(P>0.05).In the ICI182780-treated NaAsO₂ groups,the ERα,ERβ,c-MYC,Bcl-2,p-PI3K,and p-AKT protein expression increased(P<0.001,P<0.01),the Bax expression decreased(P<0.001),and the PI3K and AKT levels remained unchanged(P>0.05)compared with the corresponding NaAsO₂-only groups.The mRNA expression patterns of ERα,ERβ,c-MYC,Bcl-2,and Bax were consistent with the WB results. [Conclusion]NaAsO₂ inhibits proliferation and promotes apoptosis in Nthy-ori3-1 cells in a dose-dependent manner.The underlying mechanism likely involves NaAsO₂-mediated suppression of the ER-PI3K/AKT signaling pathway,which subsequently regulates downstream proliferation-and apoptosis-related genes.关键词
亚砷酸钠/雌激素样效应/雌激素受体/PI3K/AKT信号通路/增殖/凋亡Key words
sodium arsenite/estrogen effect/estrogen receptor/PI3K/AKT signaling pathway/proliferation/apoptosis分类
医药卫生引用本文复制引用
江志红,李宏昀,马晓薇,赖媛艳,吴军..亚砷酸钠通过调控ER-PI3K/AKT信号通路影响甲状腺正常细胞增殖与凋亡[J].环境与职业医学,2025,42(4):467-474,8.基金项目
新疆维吾尔自治区自然科学基金项目(2018D01C147) （2018D01C147）
