现代检验医学杂志2025，Vol.40Issue(6)：18-21,4.DOI:10.3969/j.issn.1671-7414.2025.06.004
STX4通过PI3K/AKT/mTOR信号通路参与调控卵巢癌细胞内质网-线粒体接触及促进迁移、侵袭的实验研究
Experimental Study on STX4 Participates in Regulating Endoplasmic Reticulum-Mitochondria Contact on Promoting Migration and Invasion in Ovarian Cancer Cells through the PI3K/AKT/mTOR Signaling Pathway
摘要
Abstract
Objective To investigate the role of syntaxin 4(STX4)in endoplasmic reticulum(ER)-mitochondria corcinoma in ovarian cancer(OC)cells.Methods The differences in STX4 expression in the OC cell line SKOV-3 and normal ovarian epithe-lial cells HOSEpiC were analyzed by Western blot and real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).SKOV-3 cell lines with stable knockdown of STX4 were constructed by lentiviral transfected cells,and the knockdown efficiency was verified by Western blot and qRT-PCR.The wound healing assay examined the changes in SKOV-3 migratory ability after STX4 knockdown,the Transwell assay examined the changes in SKOV-3 invasive ability after STX4 knockdown.The ER-mito-chondrial contact structure changes in SKOV-3 cells were visualized by transmission electron microscopy.Intracellular phospha-tidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)pathway proteins and their phosphor-ylated proteins were measured at the protein level.Results Both Western blot and qRT-PCR results showed that the expression level of STX4 was significantly higher in SKOV-3 cells than in HOSEpiC cells,and the differences were statistically significant(t=7.86,9.27,all P<0.05).The knockdown efficiency was verified by Western blot and qRT-PCR.Subsequently,the results of the wound-healing assay indicated that the migratory ability of SKOV-3 cells was significantly inhibited upon STX4 knockdown.The results of the Transwell test showed that the invasive capacity of SKOV-3 cells was significantly reduced after STX4 knock-down(t=7.56,8.82,all P<0.05).Endoplasmic reticulum-mitochondrial contact structure was reduced in SKOV-3 cells with STX4 knockdown under TEM.Finally,no significant changes after STX4 knockdown in the overall levels of PI3K/AKT/mTOR pathway proteins in SKOV-3 cells(t=2.77,5.58,2.99,all P>0.05),but their phosphorylated proteins were significantlydecreased(t=4.42,5.01,5.58,all P>0.05).Conclusion STX4 promoted OC cell migration,invasion,and the formation of ER-mitochondrial contact structures through the activation of the PI3K/AKT/mTOR pathway through phosphorylation.关键词
突触融合蛋白4/卵巢癌/内质网-线粒体接触Key words
syntaxin 4/ovarian cancer/endoplasmic reticulum-mitochondrial contacts分类
医药卫生引用本文复制引用
刘雪,卢静静,宋娟丽..STX4通过PI3K/AKT/mTOR信号通路参与调控卵巢癌细胞内质网-线粒体接触及促进迁移、侵袭的实验研究[J].现代检验医学杂志,2025,40(6):18-21,4.基金项目
河北省医学科学研究课题(课题编号:20220561). （课题编号:20220561）
