Experimental Study on the Regulatory Effects of miR-100-5p on Proliferation and Apoptosis of Thyroid Cancer Cells
10.3969/j.issn.1671-7414.2024.04.011
- VernacularTitle:miR-100-5p对甲状腺癌细胞增殖与凋亡调控作用的实验研究
- Author:
Tinghua ZHANG
1
;
Youyuan HU
;
Bo YUAN
Author Information
1. 怀化市第二人民医院检验科,湖南怀化 418000
- Keywords:
microRNA-100-5p;
thyroid cancer;
cell proliferation;
cell apoptosis
- From:
Journal of Modern Laboratory Medicine
2024;39(4):56-62
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the expression of microRNA(miR)-100-5p in thyroid cancer cells and its regulatory effects on cell proliferation and apoptosis through experiments.Methods The relative expressions of miR-100-5p in thyroid cancer cell lines(TPC-1 and KTC-1)and normal thyroid cell lines(Nthy ori3-1)were detected using fluorescence quantitative PCR.After transfection of miR-100-5p mimic,miR-100-5p inhibitor,and corresponding negative controls(miR-mimic NC,miR-inhibitor NC)into TPC-1 cells,the proliferation condition of TPC-1 cells was detected using CCK-8,and the apoptosis condition of TPC-1 cells was detected using flow cytometry.Prediction and functional enrichment analysis of target genes of miR-100-5p were performed using the miRTarBase and TargetScan7.2 databases.The targeted regulatory effect of miR-100-5p on fibroblast growth factor receptor 3(FGFR3)was validated using Western blot and dual luciferase reporter gene experiments.Results Compared with Nthy-ori3-1 cells,the expression levels of miR-100-5p in TPC-1 cells(1.87±0.03 vs 1.00±0.03)and KTC-1 cells(6.33±0.47 vs 1.00±0.03)were both up-regulated,with significant differences(t=-34.220,-19.588,all P＜0.05).The 450nm absorbance(A450nm)of cells transfected with miR-100-5p mimic at 24,48 and 72 h were higher than the miR-mimic NC group,with significant differences(t=-7.516,-17.828,-8.445,all P＜0.05).Conversely,the A450nm values of cells transfected with miR-100-5p inhibitor at 24,48 and 72 h were lower than the miR-inhibitor NC group,with significant differences(t=6.720,6.782,6.073,all P＜0.05).The apoptosis rate after transfection with miR-100-5p mimic was decreased compared to miR-mimic NC group(7.43％±0.49％vs 10.55％±0.80％),with significant differences(t=5.767,P=0.004).Compared to miR-inhibitor NC group,the apoptosis rate after transfection with miR-100-5p inhibitor was increased(3.19％±0.22％vs 2.64％±0.15％),with significant differences(t=-3.606,P=0.023).Western blot experiments showed that FGFR3 protein expression levels in the miR-100-5p mimic group were down-regulated compared to the miR-mimic NC group(0.78±0.12 vs 1.00±0.00),with significant differences(t=3.071,P=0.037).Compared to the miR-inhibitor NC group,FGFR3 protein expression levels in the miR-100-5p inhibitor group were up-regulated(1.17±0.07 vs 1.00±0.00),with significant differences(t=-4.509,P=0.046).There was no significant difference in the luciferase activity of the FGFR3 wild-type(1.01±0.17 vs 1.00±0.00)and mutant groups(0.99±0.11 vs 1.00±0.00)between miR-100-5p mimic and miR-mimic NC,and the differences were statistically significant(f=-0.057,0.181,P=0.96,0.873).Conclusion MiR-100-5p in thyroid cancer cells was up-regulated,which may promote cell proliferation and inhibit apoptosis.It may become a new biomarker and regulatory target in the diagnosis and treatment of thyroid cancer.
