Abstract
Multiple circular RNAs (circRNAs) have been identified to act as essential mediators in diverse human cancers. However, the roles of circRNAs in neuroblastoma (NB) are largely unknown. In this study, we aimed to explore the function of circKIF2A in NB. Quantitative real-time polymerase chain reaction was executed to detect the levels of circKIF2A, KIF2A mRNA, miR-129-5p and polo-like kinase 4 (PLK4) mRNA. Actinomycin D assay and RNase R digestion assay were conducted to analyze the feature of circKIF2A. 3-(4, 5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, transwell assay and specific kits were utilized to evaluate cell proliferation, metastasis and glycolysis, respectively. Western blot assay was performed to examine the protein levels of matrix metalloproteinase 2 (MMP2), MMP9 and PLK4. Bioinformatics analysis, RNA pull-down assay and dual-luciferase reporter assay were conducted to analyze the relationship between miR-129-5p and circKIF2A or PLK4. Murine xenograft model assay was done to investigate the role of circKIF2A in NB in vivo. CircKIF2A level was increased in NB tissue samples and cell lines. Silencing of circKIF2A impeded NB cell proliferation, migration, invasion and glycolysis. For mechanism analysis, circKIF2A could positively modulate PLK4 expression via sponging miR-129-5p. Moreover, miR-129-5p inhibition reversed the inhibitory effects of circKIF2A silencing on the behaviors of NB cells. MiR-129-5p overexpression weakened the malignant biological behaviors of NB cells by targeting PLK4. Additionally, circKIF2A knockdown hampered tumorigenesis in vivo. CircKIF2A knockdown suppressed cell proliferation, migration, invasion and glycolysis via downregulating PLK4 expression through miR-129-5p.
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Abbreviations
- NB:
-
Neuroblastoma
- PLK4:
-
Polo-like kinase 4
- MMP2:
-
Matrix metalloproteinase 2
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11010_2021_4096_MOESM1_ESM.tif
Figure S1. The selection of miR-129-5p and PLK4. (A) The expression levels of (miR-367-5p, miR-6134, miR-6839-3p, miR-6851-5p, miR-1250-3p, miR-1270, miR-129-5p, miR-1301-3p, miR-136-5p and miR-154-3p in NB tissues were detected by qRT-PCR assay. (B and C) The expression levels of miR-129-5p and miR-1301-3p in SK-N-SH and SK-N-AS cells transfected with si-NC or si-circKIF2A were detected by qRT-PCR assay. (D) The levels of SIAH1, PLK1, BCL2, HOXA7, CREB1 and BCL11A in NB tissues were measured by qRT-PCR assay. (E and F) The levels of SIAH1, PLK1, BCL2, HOXA7, CREB1 and BCL11A in SK-N-SH and SK-N-AS cells transfected with miR-NC or miR-129-5p were detected by qRT-PCR assay. *P<0.05. Supplementary file1 (TIF 759 KB)
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Yang, Y., Pan, H., Chen, J. et al. CircKIF2A contributes to cell proliferation, migration, invasion and glycolysis in human neuroblastoma by regulating miR-129-5p/PLK4 axis. Mol Cell Biochem 476, 2513–2525 (2021). https://doi.org/10.1007/s11010-021-04096-3
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DOI: https://doi.org/10.1007/s11010-021-04096-3