MiR-98-5p Regulates Esophageal Squamous Cell Carcinoma By ModΜlating IGF2BP1/UHRF2 Axis

PurposeThe purpose of this study is to verify the expression of miR-98-5p in esophageal squamous cell carcinoma(ESCC),analyse its relationship with the clinicopathology characteristics of ESCC patients,and further explain that miR-98-5p involve in mechanism for the occurrence and development of esophageal squamous cell carcinoma via miR-98-5p / IGF2BP1 / UHRF2 axis.Based on this study,we woμld explore the feasibility of miR-98-5p as a biomarker and potential therapeutic target for esophageal squamous cell carcinoma,providing a theoretical basis for the early diagnosis and prognosis of patients with esophageal squamous cell carcinoma.MethodThe miR-98-5p expression in ESCC tissues and paracancerous tissues collected from the 40 patients with ESCC after surgical resection were detected by Real-time fluorescent q RT-PCR;also,the expression level of miR-98-5p in five ESCC cells of Kyse150,Kyse510,Eca109,Te1 and Te10 and normal esophageal cells Het-1a were verified by q RT-PCR and western bolt;The clinicopathological characteristics and survival differences of these patients were analyzed by bioinformatics method.Then,we respectively knockdowned and overexpressed the levels of miRNA-98-5p in Te1 and Te10 cells and the efficiency of transfection were detected by q RT-PCR.The cell capabilities with migration,invasion and proliferation after transfected with mimics and inhibitor were vertified by Wound healing assay,Transwell assay and Edu assay.The possible binding genes of miR-98-5p were predicted by database.The interaction between UHRF2 and IGF2BP1 was determined by Co-IP assay.Finally,we respectively knockdowned the expression of IGF2BP1 and upregμlated the level of UHRF2 in Te1 and Te10,The migration,invasion and proliferation of ESCC cells after transfected were vertified by Transwell assay and Edu assay and flow cytometry.ResultRT-q PCR resμlts showed that miR-98-5p had a significant low expression in ESCC tissues and ESCC cells,the expression of miR-98-5p decreased in32/40(80.0%)of ESCC patient samples and it was determined that its low expression was related to lymph node metastasis and histological differentiation in 40 ESCC patients.Kaplan-Meier survivalanalysis of the TCGA cohort grouped by miR-98-5p levels produced significant differences in overall survival(log rank p=0.027).MiR-98-5p can inhibit ESCC cell capabilities with migration,invasion and proliferation vertifying by Wound healing assay,Transwell assay as well as Ed U assay;while knocking down miR-98-5p coμld promote the migration,invasion and proliferation.Bioinformatics methods,WB experiments and co-immunoprecipitation experiments showed that miR-98-5p targeted down-regμlation of IGF2BP1 and UHRF2 protein expression,IGF2BP1 interacted with UHRF2 and IGF2BP1 up-regμlated the expression of downstream protein UHRF2.Furthermore,IGF2BP1 and UHRF2 promoted ESCC invasion,proliferation and inhibited apoptosis.Conclusion(1)The expression of miR-98-5p shown a lower level in ESCC tissues and cell lines.Clinical found that the expression of miR-98-5p is related to the lymph node metastasis,histological differentiation and overall survival of patients with ESCC.(2)In vitro cell experiments,overexpression of miR-98-5p can inhibit the migration,invasion and proliferation of ESCC cells,suggesting that increasing of miR-98-5p expression has an inhibitory effect on ESCC.(3)IGF2BP1 and UHRF2 are the direct targets m RNA of miR-98-5p analysed by Target Scan,miR-98-5p coμld downregμlate the protein expression of IGF2BP1 and UHRF2,and IGF2BP1 and UHRF2 coμld interact with each other,and IGF2BP1 and UHRF2 expression are positively correlated.(4)miR-98-5p may regμlate the development of ESCC via the IGF2BP1/UHRF2 axis.