NUDT5 promotes the growth, metastasis, and Warburg effect of IDH wild-type glioblastoma multiforme cells by upregulating TRIM47

Abstract

Objective: To explore the regulatory mechanism of NUDT5 in glioblastoma multiforme (GBM). Methods: GEPIA database was used to predict the expressions of NUDT5 and tripartite motif family proteins 47 (TRIM47) in GBM patients. RT-qPCR and Western blot analyses were performed to examine NUDT5 expression in GBM cells. LN-229 cell proliferation, migration as well as invasion were estimated by CCK-8, colony formation, wound healing, and Transwell assays following interference with NUDT5. ECAR assay, L-lactic acid kit, glucose detection kit, and ATP detection kit were applied for the detection of glycolysis-related indexes. Co-immunoprecipitation experiment was carried out to verify the relationship between NUDT5 and TRIM47. Results: GEPIA database showed that NUDT5 expression was significantly increased in GBM patients. Inhibiting the expression of NUDT5 in GBM cells significantly suppressed the viability, proliferation, invasion, migration, and glycolysis of GBM cells. Moreover, TRIM47 was highly expressed in GBM cells and interacted with NUDT5. Overexpression of TRIM47 partially reversed the inhibitory effect of NUDT5 downregulation on the proliferation, metastasis, and glycolysis of GBM cells. Conclusions: NUDT5 promotes the growth, metastasis, and Warburg effect of GBM cells by upregulating TRIM47. Both NUDT5 and TRIM47 can be used as targets for GMB treatment.

Keywords

Glioblastoma multiforme / NUDT5 / TRIM47 / Growth / Metastasis / Warburg effect

Conflict of interest statement

The authors declare that they have no conflict of interest.

Funding

The authors received no extramural funding for this study.

Data availability statement

The data supporting the findings of this study are available from the corresponding authors upon request.

Authors’ contributions

ZFZ and SML conceived and designed the study. ZFZ carried out the experiments. SML analyzed the experimental data. ZFZ and SML wrote and revised the manuscript and confirmed the authenticity of all the raw data. All authors have read and approved the final manuscript.

References

[1] Incekara F, Smits M, van der Voort SR, Dubbink HJ, Atmodimedjo PN, Kros JM, et al. The association between the extent of glioblastoma resection and survival in light of MGMT promoter methylation in 326 patients with newly diagnosed IDH-wildtype glioblastoma. Front Oncol 2020; 10: 1087.

[2] Tirro E, Massimino M, Broggi G, Romano C, Minasi S, Gianno F, et al. A custom DNA-based NGS panel for the molecular characterization of patients with diffuse gliomas: Diagnostic and therapeutic applications. Front Oncol 2022; 12: 861078.

[3] Caltabiano R, Torrisi A, Condorelli D, Albanese V, Lanzafame S. High levels of connexin 43 mRNA in high grade astrocytomas. Study of 32 cases with in situ hybridization. Acta Histochem 2010; 112(6): 529-535.

[4] Patil CG, Yi A, Elramsisy A, Hu J, Mukherjee D, Irvin DK, et al. Prognosis of patients with multifocal glioblastoma: A case-control study. J Neurosurg 2012; 117(4): 705-711.

[5] Wang Y, Wan F, Chang K, Lu X, Dai B, Ye D. NUDT expression is predictive of prognosis in patients with clear cell renal cell carcinoma. Oncol Lett 2017; 14(5): 6121-6128.

[6] Zhang L, Song XN, Dai DP, Zhou XY, Gan W, Takagi Y, et al. Lowered Nudix type 5 expression leads to cellular senescence in IMR-90 fibroblast cells. Free Radic Res 2013; 47(6-7): 511-516.

[7] Zhang LQ, Dai DP, Gan W, Takagi Y, Hayakawa H, Sekiguchi M, et al. Lowered nudix type 5 (NUDT5) expression leads to cell cycle retardation in HeLa cells. Mol Cell Biochem 2012; 363(1-2): 377-384.

[8] Wang JJ, Liu TH, Li J, Li DN, Tian XY, Ouyang QG, et al. The high expression of MTH1 and NUDT5 predict a poor survival and are associated with malignancy of esophageal squamous cell carcinoma. Peer J 2020; 8: e9195. doi: 10.7717/peerj.9195.

[9] Tian Y, Chen L, Jiang Y. LASSO-based screening for potential prognostic biomarkers associated with glioblastoma. Front Oncol 2022; 12: 1057383.

[10] Fujimura T, Inoue S, Urano T, Takayama K, Yamada Y, Ikeda K, et al. Increased expression of tripartite motif (TRIM) 47 is a negative prognostic predictor in human prostate cancer. Clin Genitourin Cancer 2016; 14(4): 298-303.

[11] Li W, Zhang W, Deng W, Zhong Y, Zhang Y, Peng Z, et al. Quantitative proteomic analysis of mitochondrial proteins differentially expressed between small cell lung cancer cells and normal human bronchial epithelial cells. Thorac Cancer 2018; 9(11): 1366-1375.

[12] Liang Q, Tang C, Tang M, Zhang Q, Gao Y, Ge Z. TRIM47 is upregulated in colorectal cancer, promoting ubiquitination and degradation of SMAD4. J Exp Clin Cancer Res 2019; 38(1): 159.

[13] Chen L, Li M, Li Q, Xu M, Zhong W. Knockdown of TRIM47 inhibits glioma cell proliferation, migration and invasion through the inactivation of Wnt/beta-catenin pathway. Mol Cell Probes 2020; 53: 101623.

[14] Li L, Yu Y, Zhang Z, Guo Y, Yin T, Wu H, et al. TRIM47 accelerates aerobic glycolysis and tumor progression through regulating ubiquitination of FBP1 in pancreatic cancer. Pharmacol Res 2021; 166: 105429.

[15] Wang Y, Liu C, Xie Z, Lu H. Knockdown of TRIM47 inhibits breast cancer tumorigenesis and progression through the inactivation of PI3K/Akt pathway. Chem Biol Interact 2020; 317: 108960.

[16] Li C, Tang Z, Zhang W, Ye Z, Liu F. GEPIA2021: Integrating multiple deconvolution-based analysis into GEPIA. Nucleic Acids Res 2021; 49(W1): W242-W246.

[17] Warde-Farley D, Donaldson SL, Comes O, Zuberi K, Badrawi R, Chao P, et al. The GeneMANIA prediction server: Biological network integration for gene prioritization and predicting gene function. Nucleic Acids Res 2010; 38(Web Server issue): W214-W220.

[18] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001; 25(4): 402-408.

[19] Ohgaki H, Kleihues P. Population-based studies on incidence, survival rates, and genetic alterations in astrocytic and oligodendroglial gliomas. J Neuropathol Exp Neurol 2005; 64(6): 479-489.

[20] Wang Q, Lu X, Wang J, Yang Z, Hoffman RM, Wu X. Moesin upregulation is associated with enhanced tumor progression imaged noninvasively in an orthotopic mouse model of human glioblastoma. Anticancer Res 2018; 38(6): 3267-3272.

[21] Zhang H, Zhang LQ, Yang CC, Li J, Tian XY, Li DN, et al. The high expression of NUDT5 indicates poor prognosis of breast cancer by modulating AKT/Cyclin D signaling. PLoS One 2021; 16(2): e0245876.

[22] Li DN, Yang CC, Li J, Ouyang QG, Zeng LT, Fan GQ, et al. The high expression of MTH1 and NUDT5 promotes tumor metastasis and indicates a poor prognosis in patients with non-small-cell lung cancer. Biochim Biophys Acta Mol Cell Res 2021; 1868(1): 118895.

[23] Liberti MV, Locasale JW. The Warburg effect: How does it benefit cancer cells? Trends Biochem Sci 2016; 41(3): 211-218.

[24] Wei H, Ding C, Zhuang H, Hu W. TRIM47 promotes the development of glioma by ubiquitination and degradation of FOXO1. Onco Targets Ther 2020; 13: 13401-13411.

[25] Xia Y, Wei Z, Huang W, Wei X, He Y. Trim47 overexpression correlates with poor prognosis in gastric cancer. Neoplasma 2021; 68(2): 307-316.