Effects of insulin-like growth factor 2mRNA-binding protein 3 on the malignant phenotype of esophageal cancer cells
CHANG Huiwen
SHI Woda
SHI Zhan
ZHANG Yajun
Abstract：Objective To investigate the effects of insulin-like growth factor 2mRNA-binding protein 3(IGF2BP3)on the malignant phenotype of esophageal cancer cells and its upstream regulatory mechanisms.Methods Bioinformatics analysis combined with real-time quantitative PCR and Western blot were employed to detect the expression of IGF2BP3 and Tet methylcytosine dioxygenase1(TET1),while the correlation between IGF2BP3 and TET1 expression was analyzed in public databases.In esophageal cancer cells TE-15 and ESC-410 transfected with IGF2BP3-specific small interference RNA(siRNA;si-IGF2BP3 group)and negative control siRNA(si-NC group),cell viability,cell proliferation ability and migration and invasion capacities were assessed by Cell Counting Kit-8,5-ethynyl-2'-deoxyuridine(EdU)and Transwell migration and Matrigel invasion assays,and apoptosis was determined by flow cytometry.Western blot was performed to detect the protein expression levels of Ki67,matrix metalloproteinase(MMP)-7/9,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and cleaved caspase-3(CC3).In cells transfected with TET1 overexpression plasmid(Oe-TET1 group)and empty vector plasmid(Oe-NC group),or TET1-specific siRNA(si-TET1 group)and si-NC group,Western blot and RNA methylated immunoprecipitation were utilized to evaluate the protein expression level of IGF2BP3 and the N6-methyladenosine(m6A)methylation degree.Results Compared to normal tissues,the expression of both IGF2BP3 and TET1 was significantly up-regulated in esophageal cancer tissues(P<0.001),and a significant positive correlation was observed between their expression levels(r=0.220,P<0.001).Experimental validation demonstrated that in esophageal cancer cell lines(Eca-109,TE-1,EC-1,ESC-410),the mRNA and protein expression levels of IGF2BP3 and TET1 were higher than those in the normal human esophageal epithelial cell line HET-1A,with the highest expression observed in TE-15 and ESC-410 cells(P<0.001).In functional studies,results showed that compared to the si-NC group,the si-IGF2BP3 group exhibited significantly reduced proliferative activity,decreased EdU-positive rate,weakened migration and invasion abilities,and a significantly increased apoptosis rate(P<0.001).The protein expression levels of Ki-67,MMP-7,MMP-9,and Bcl-2 were down-regulated in the si-IGF2BP3 group,while the expression levels of Bax and CC3 were up-regulated(P<0.001).In mechanistic studies,compared to the Oe-NC group,the Oe-TET1 group showed significantly increased m6A methylation level and protein expression of IGF2BP3;whereas compared to the si-NC group,the si-TET1 group exhibited significantly reduced m6A methylation level and protein expression of IGF2BP3(P<0.001).Conclusion In esophageal cancer,TET1 potentially upregulates IGF2BP3 by enhancing its m⁶A methylation,thereby driving malignant progression.Consequently,the TET1/IGF2BP3 axis represents a promising potential therapeutic target.
Keywords：Esophageal cancerInsulin-like growth factor 2mRNA-binding protein 3Tet methylcytosine dioxygenase1Malignant phenotype
Publication Date：2025-10-28
Online Publishing Date：2025-12-03(First online date of this platform, not the publication date of the document)
Pages：7( 937-943 )