Anti-Migratory Effect of Dipotassium Glycyrrhizinate on Glioblastoma Cell Lines: Microarray Data for the Identification of Key MicroRNA Signatures

Overview

Journal Front Oncol

Specialty Oncology

Date 2022 Aug 22

PMID 35992881

Authors

Gabriel Alves Bonafe

Jessica Silva Dos Santos

Anna Maria Alves de Piloto Fernandes

Jussara Vaz Ziegler

Fernando Augusto Lima Marson

Thalita Rocha

Patricia de Oliveira Carvalho

Manoela Marques Ortega

Affiliations

Soon will be listed here.

Abstract

The nuclear factor kappa B (NF-κB) pathway has been reported to be responsible for the aggressive disease phenomenon observed in glioblastoma (GBM). Dipotassium glycyrrhizinate (DPG), a dipotassium salt of glycyrrhizic acid isolated from licorice, has recently demonstrated an anti-tumoral effect on GBM cell lines U87MG and T98G through NF-κB suppression by - and -mediating microRNA (miR)-16 and miR-146a, respectively. Thus, the present study aimed to evaluate the expression profiles of miRNAs related to NF-κB suppression in T98G GBM cell line after DPG exposure using miRNA microarray (Affymetrix Human miRNA 4.0A), considering only predicted miRNAs as NF-κB regulator genes. Additional assays using U251 and U138MG cells were performed to validate the array results. DPG cytotoxicity was determined by (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, and cellular apoptosis was quantified by DNA fragmentation and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. The anti-proliferative effect was observed by cell proliferation and wound-healing assays, and the sphere formation assay examined whether DPG reduced stem cell subpopulation formation. The most over-expressed miRNAs were miR-4443 and miR-3620. The cytotoxic effect of DPG in U251 and U138MG was observed with an IC50 of 32 and 20 mM for 48 h, respectively. The IC50 of each cell line was used in all further assays. DPG treatment-induced apoptosis is observed by DNA fragmentation and increased TUNEL-positive cells. Cell proliferation and wound-healing assays showed an anti-proliferative and anti-migratory effect by DPG on the evaluated cell lines. In addition, DPG treatment led to a 100% reduction in sphere formation. The qPCR results in U251 and U138MG cells showed that DPG increased miR-4443 (2.44 . 1.11, -value = 0.11; 8.27 . 1.25, -value = 0.04) and miR-3620 expression (1.66 vs. 1.00, -value = 0.03; 8.47 . 1.01, -value = 0.03) and decreased (0.44 . 1.10, -value = 0.03; 0.49 . 1.07, -value = 0.04) and (0.20 . 1.03, -value = 0.001; 0.39 . 1.06, -value = 0.01) mRNA levels compared to controls. Our results suggest that DPG inhibits cell viability by activating apoptosis and inhibiting cell proliferation and stem cell subpopulation formation through miR-4443 and miR-3620 upregulation. Both miRNAs are responsible for the post-transcriptional inhibition of NF-κB by and modulation.

Citing Articles

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The Recent Research Progress of NF-κB Signaling on the Proliferation, Migration, Invasion, Immune Escape and Drug Resistance of Glioblastoma.

Shi P, Xu J, Cui H Int J Mol Sci. 2023; 24(12).

PMID: 37373484 PMC: 10298967. DOI: 10.3390/ijms241210337.

References

Verhaak R, Hoadley K, Purdom E, Wang V, Qi Y, Wilkerson M . Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1. Cancer Cell. 2010; 17(1):98-110. PMC: 2818769. DOI: 10.1016/j.ccr.2009.12.020. View

Yang G, Han D, Chen X, Zhang D, Wang L, Shi C . MiR-196a exerts its oncogenic effect in glioblastoma multiforme by inhibition of IκBα both in vitro and in vivo. Neuro Oncol. 2014; 16(5):652-61. PMC: 3984554. DOI: 10.1093/neuonc/not307. View

Huang S, Ali N, Zhong L, Shi J . MicroRNAs as biomarkers for human glioblastoma: progress and potential. Acta Pharmacol Sin. 2018; 39(9):1405-1413. PMC: 6289388. DOI: 10.1038/aps.2017.173. View

Sottoriva A, Spiteri I, Piccirillo S, Touloumis A, Collins V, Marioni J . Intratumor heterogeneity in human glioblastoma reflects cancer evolutionary dynamics. Proc Natl Acad Sci U S A. 2013; 110(10):4009-14. PMC: 3593922. DOI: 10.1073/pnas.1219747110. View

Brennan C, Verhaak R, McKenna A, Campos B, Noushmehr H, Salama S . The somatic genomic landscape of glioblastoma. Cell. 2013; 155(2):462-77. PMC: 3910500. DOI: 10.1016/j.cell.2013.09.034. View

Ebrahimi S, Reiisi S . Downregulation of miR-4443 and miR-5195-3p in ovarian cancer tissue contributes to metastasis and tumorigenesis. Arch Gynecol Obstet. 2019; 299(5):1453-1458. DOI: 10.1007/s00404-019-05107-x. View

Wu H, Liu Q, Cai T, Chen Y, Wang Z . Induction of microRNA-146a is involved in curcumin-mediated enhancement of temozolomide cytotoxicity against human glioblastoma. Mol Med Rep. 2015; 12(4):5461-6. DOI: 10.3892/mmr.2015.4087. View

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

. Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature. 2008; 455(7216):1061-8. PMC: 2671642. DOI: 10.1038/nature07385. View

10.

Annovazzi L, Caldera V, Mellai M, Riganti C, Battaglia L, Chirio D . The DNA damage/repair cascade in glioblastoma cell lines after chemotherapeutic agent treatment. Int J Oncol. 2015; 46(6):2299-308. PMC: 4441296. DOI: 10.3892/ijo.2015.2963. View

11.

Stupp R, Mason W, van den Bent M, Weller M, Fisher B, Taphoorn M . Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005; 352(10):987-96. DOI: 10.1056/NEJMoa043330. View

12.

Yang T, Lu X, Wu T, Ding D, Zhao Z, Chen G . MicroRNA-16 inhibits glioma cell growth and invasion through suppression of BCL2 and the nuclear factor-κB1/MMP9 signaling pathway. Cancer Sci. 2014; 105(3):265-71. PMC: 4317940. DOI: 10.1111/cas.12351. View

13.

Rajbhandari R, McFarland B, Patel A, Gerigk M, Gray G, Fehling S . Loss of tumor suppressive microRNA-31 enhances TRADD/NF-κB signaling in glioblastoma. Oncotarget. 2015; 6(19):17805-16. PMC: 4627347. DOI: 10.18632/oncotarget.4596. View

14.

Snuderl M, Fazlollahi L, Le L, Nitta M, Zhelyazkova B, Davidson C . Mosaic amplification of multiple receptor tyrosine kinase genes in glioblastoma. Cancer Cell. 2011; 20(6):810-7. DOI: 10.1016/j.ccr.2011.11.005. View

15.

Gao Y, Xu Y, Wang J, Yang X, Wen L, Feng J . lncRNA MNX1-AS1 Promotes Glioblastoma Progression Through Inhibition of miR-4443. Oncol Res. 2018; 27(3):341-347. PMC: 7848264. DOI: 10.3727/096504018X15228909735079. View

16.

Bonafe G, Dos Santos J, Ziegler J, Umezawa K, Ribeiro M, Rocha T . Growth Inhibitory Effects of Dipotassium Glycyrrhizinate in Glioblastoma Cell Lines by Targeting MicroRNAs Through the NF-κB Signaling Pathway. Front Cell Neurosci. 2019; 13:216. PMC: 6546822. DOI: 10.3389/fncel.2019.00216. View

17.

Galardi S, Mercatelli N, Farace M, Ciafre S . NF-kB and c-Jun induce the expression of the oncogenic miR-221 and miR-222 in prostate carcinoma and glioblastoma cells. Nucleic Acids Res. 2011; 39(9):3892-902. PMC: 3089483. DOI: 10.1093/nar/gkr006. View

18.

Brassesco M, Roberto G, Morales A, Oliveira J, Delsin L, Pezuk J . Inhibition of NF- κ B by Dehydroxymethylepoxyquinomicin Suppresses Invasion and Synergistically Potentiates Temozolomide and γ -Radiation Cytotoxicity in Glioblastoma Cells. Chemother Res Pract. 2013; 2013:593020. PMC: 3594939. DOI: 10.1155/2013/593020. View

19.

Kim T, Huang W, Park R, Park P, Johnson M . A developmental taxonomy of glioblastoma defined and maintained by MicroRNAs. Cancer Res. 2011; 71(9):3387-99. PMC: 3085663. DOI: 10.1158/0008-5472.CAN-10-4117. View

20.

Hambardzumyan D, Bergers G . Glioblastoma: Defining Tumor Niches. Trends Cancer. 2016; 1(4):252-265. PMC: 4831073. DOI: 10.1016/j.trecan.2015.10.009. View