中国病理生理杂志2024，Vol.40Issue(7)：1190-1196,7.DOI:10.3969/j.issn.1000-4718.2024.07.005
ANKRD49通过增加Snail/Slug/ZEB1表达促进NCI-H1299细胞上皮-间充质转化的研究
ANKRD49 promoted epithelial-mesenchymal transition by increasing ex-pression of Snail/Slug/ZEB1 in NCI-H1299 cells
摘要
Abstract
AIM:To investigate the effect of ankyrin repeat domain 49(ANKRD49)on epithelial-mesenchy-mal transition(EMT)in NCI-H1299 cells,and to explore its mechanism.METHODS:The ANKRD49 was over-ex-pressed in NCI-H1299 cells.The morphological changes of ANKRD49-overpressing NCI-H1299 cells were observed under microscope.The mRNA and protein expression levels of EMT-related markers[E-cadherin,transforming growth factor-β1(TGF-β1),vimentin and α-smooth muscle actin(α-SMA)]and EMT-related transcription factors(Snail1,Slug,Twist and ZEB1)were detected by RT-qPCR Western blot.Immunofluorescence staining was performed to observe the localiza-tion and expression of E-cadherin and vimentin in ANKRD49-overexpressing cells or control cells.Immunohistochemical method was performed to examine the levels of E-cadherin,α-SMA,Snail,Slug and ZEB1 in lung tissues of nude mice in-oculated with ANKRD49-overexpressing H1299 cells or control cells.RESULTS:Compared with the control group,the ANKRD49 overexpressing cells showed mesenchymal cell morphology(fusiform and less tight connections).RT-qPCR and Western blot results showed that the mRNA and protein levels of mesenchymal markers vimentin and α-SMA in ANKRD49 overexpressing cells were significantly higher than those in cells of control group,while the mRNA and protein levels of epithelial marker E-cadherin were lower than those in cells of control group.Compared with control group,the im-munofluorescence intensity of E-cadherin of H1299 cells decreased in after ANKRD49 overexpression,while that of vimen-tin increased significantly.Snail,Slug and ZEB1 expression were significantly elevated in ANKRD49 overexpressing cells compared with control group.The levels of E-cadherin in lung tissues of nude mice inoculated with ANKRD49-overexpressing H1299 cells declined,while the levels of α-SMA,Snail,Slug and ZEB1 increased compared with those in control mice.CONCLUSION:ANKRD49 promoted EMT of NCI-H1299 cells by increasing the expression of Snail1,Slug and ZEB1 and consequent downreguation of E-cadherin and upregulation of vimentin and α-SMA.关键词
锚蛋白重复结构域蛋白49/肺腺癌/上皮-间充质转化Key words
ankyrin repeat domain 49/lung adenocarcinoma/epithelial-mesenchymal transition分类
医药卫生引用本文复制引用
高睿,刘超锋,胡金瑞,梁刚,付荣,王维,王海龙,庞敏..ANKRD49通过增加Snail/Slug/ZEB1表达促进NCI-H1299细胞上皮-间充质转化的研究[J].中国病理生理杂志,2024,40(7):1190-1196,7.基金项目
山西省自然科学基金资助项目(No.202203021212041) （No.202203021212041）
