circ_100284 via miR-217/MAPK1 regulates esophageal squamous cell carcinoma invasion and its effects on 5-FU chemotherapy sensitivity
10.11855/j.issn.0577-7402.0787.2024.0201
- VernacularTitle:circ_100284通过miR-217/MAPK1调控食管鳞状细胞癌侵袭及其对5-FU化疗敏感性的影响
- Author:
Yan-Wei SHI
1
;
Cai-Feng MI
;
Li-Lin NIU
;
Yang YANG
Author Information
1. 平顶山学院第一附属医院/平顶山市第一人民医院消化内科,河南 平顶山 467000
- Keywords:
circ_100284;
miR-217;
mitogen-activated protein kinase 1;
esophageal cancer;
chemosensitivity
- From:
Medical Journal of Chinese People's Liberation Army
2024;49(10):1184-1195
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of circ_100284 in affecting the invasion of esophageal squamous cell carcinoma(ESCC)cells and their sensitivity to 5-fluorouracil(5-FU)chemotherapy via regulating miR-217/mitogen-activated protein kinase 1(MAPK1)acting as a competing endogenous RNA(ceRNA).Methods The bioinformatics approach was used to analyze the differentially expressed circRNAs in ESCC.qRT-PCR was used to detect the expression of circ_100284 in ESCC tissues and cells.The 5-FU-resistant KYSE450 cell line(KYSE45-R)was established by increasing concentrations of 5-FU.The IC50 of 5-FU in KYSE450 and KYSE450-R cells was determined through MTT assay.qRT-PCR was used to detect the expression of circ_100284,miR-217,and MAPK1 mRNA in KYSE450 and KYSE450-R cells,while Western blotting detecting the protein expression of MAPK1.In the experiments with KYSE450-R cells,we set up the following groups:(1)blank group(without treatment),si-NC group(transfected with si-NC),si-circ_100284 group(transfected with si-circ_100284),pc-Control group(transfected with pc-Control),pc-circ_100284 group(transfected with pc-circ_100284),pc-circ_100284+mimic NC group(transfected with pc-circ_100284 and mimic NC),and pc-circ_100284+miR-217 mimic group(transfected with pc-circ_100284 and miR-217 mimic).These groups were subjected to MTT assay to detect cell viability,Transwell assay to detect cell invasion,and flow cytometry to detect cell apoptosis.(2)Blank group(without treatment),si-NC group(transfected with si-MAPK1 negative control),si-MAPK1 group(transfected with si-MAPK1),si-MAPK1+inhibitor NC group(transfected with si-MAPK1 and inhibitor NC),and si-MAPK1+miR-217 inhibitor group(transfected with si-MAPK1 and miR-217 inhibitor).We detected the mRNA and protein expression of MAPK1 using qRT-PCR and Western blotting.We evaluated cell viability using MTT assay,invasion with Transwell assay,and apoptosis by flow cytometry.circ_100284-WT or circ_100284-MUT reporter plasmids,as well as MAPK1-WT or MAPK1-MUT reporter plasmids,were co-transfected with miR-NC or miR-217 mimic into KYSE450-R cells for 48 h,and dual luciferase reporter assay was used to measure luciferase activity.Results The bioinformatics analysis revealed significant upregulation of circ_100284 in ESCC.Compared with adjacent normal tissues,the expression of circ_100284 in ESCC tissues is enhanced(P<0.05);compared with the HECC cells,the TE-11,ECA109 and KYSE450 ESCC cell lines showed enhanced expression of circ_100284(P<0.05).Compared with the KYSE450 cells,KYSE450-R cells demonstrated increased IC50 with enhanced expression of circ_100284 and MAPK1 but suppressed expression of miR-217(P<0.05).Compared with the si-NC group,the si-circ_100284 group demonstrated inhibited invasion and proliferation of cells with increased apoptosis(P<0.05).Compared with pc-Control group,the invasion and proliferation of cells in the pc-circ_100284 group are increased,and cell apoptosis is decreased(P<0.05).Over-expression of miR-217 reversed the malignant biological behavior of ESCC cells induced by pc-circ_100284(P<0.05).Compared with si-NC group,in the si-MAPK1 group,we observed decreased cell invasion and proliferation,and increased apoptosis(P<0.05),but miR-217 inhibitor reversed the effect of si-MAPK1 on the biological behavior of ESCC cells(P<0.05).The targeting relationship of circ_100284 and miR-217,miR-217 and MAPK1 is confirmed.Conclusion circ_100284 promotes ESCC cell invasion by regulating miR-217/MAPK1,inhibits the chemosensitivity of ESCC cells to 5-FU,and acts as a tumor-promoting factor in ESCC.
