ANKRD49 promoted epithelial-mesenchymal transition by increasing ex-pression of Snail/Slug/ZEB1 in NCI-H1299 cells
10.3969/j.issn.1000-4718.2024.07.005
- VernacularTitle: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
- Author:
Rui GAO
1
;
Chaofeng LIU
;
Jinrui HU
;
Gang LIANG
;
Rong FU
;
Wei WANG
;
Hailong WANG
;
Min PANG
Author Information
1. 山西医科大学第一医院呼吸与危重症医学科,呼吸疾病防治与基础研究山西省重点实验室,山西 太原 030001
- Keywords:
ankyrin repeat domain 49;
lung adenocarcinoma;
epithelial-mesenchymal transition
- From:
Chinese Journal of Pathophysiology
2024;40(7):1190-1196
- CountryChina
- Language:Chinese
-
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.
