The impact of glutamine deprivation on the expression of MEIS3, SPAG4, LHX1, LHX2, and LHX6 genes in ERN1 knockdown U87 glioma cells

Abstract

Objective. The aim of the current study was to investigate the expression of genes encoded homeobox proteins such as MEIS3 (Meis homeobox 3), SPAG4 (sperm associated antigen 4), LHX1 (LIM homeobox 1), LHX2, and LHX6 in U87 glioma cells in response to glutamine deprivation in control glioma cells and cells with knockdown of ERN1 (endoplasmic reticulum to nucleus signaling 1), the major pathway of the endoplasmic reticulum stress signaling, for evaluation of a possible dependence on the expression of these important regulatory genes from glutamine supply and ERN1 signaling. Methods. The expression level of MEIS3, SPAG4, LHX, LHX2, and LHX6 genes was studied by real-time quantitative polymerase chain reaction in control U87 glioma cells (transfected by vector) and cells with ERN1 knockdown after exposure to glutamine deprivation. Results. It was shown that the expression level of MEIS3 and LHX1 genes was up-regulated in control glioma cells treated by glutamine deprivation. At the same time, the expression level of three other genes (LHX2, LHX6, and SPAG4) was down-regulated. Furthermore, ERN1 knockdown significantly modified the effect of glutamine deprivation on LHX1 gene expression in glioma cells, but did not change significantly the sensitivity of all other genes expression to this experimental condition. Conclusion. The results of this investigation demonstrate that the exposure of U87 glioma cells under glutamine deprivation significantly affected the expression of all genes studied encoding the homeobox proteins and that this effect of glutamine deprivation was independent of the endoplasmic reticulum stress signaling mediated by ERN1, except LHX1 gene.

ERN1 dependent impact of glutamine and glucose deprivations on the pyruvate dehydrogenase genes expression in glioma cells

“…These results clearly demonstrated that the expression of PDHA1, PDHB, DLAT, and DLD genes is controlled by glutamine supply and that this control is ER stress dependent particularly through ERN1 signaling. This is in accordance with the data our previous studies (Minchenko et al , 2020(Minchenko et al , 2021Tsymbal et al 2016;Krasnytska et al 2022) as well as studies of other authors (Drogat et al 2007). The changes in PDHA1 and PDHB genes expression under glutamine deprivation are in agreement with data of Yetkin-Arik et al (2019) indicating that silencing of PDHA1 expression prevents the proliferation of HUVECs, because this enzyme of PDH complex is a crucial regulator of pyruvate metabolic pathways (Zaher et al 2021).…”

“…Glutamine and glucose supply as well as endoplasmic reticulum (ER) stress are very important and complementary factors for the tumor growth. This is also confirmed by the fact that inhibition of endoplasmic reticulum to nucleus signaling 1 (ERN1)/inositol requiring enzyme 1 (IRE1) modifies the effect of glutamine and glucose deprivations on the expression of numerous genes (Minchenko et al , 2015a(Minchenko et al , 2020(Minchenko et al , 2021Tsymbal et al 2016Tsymbal et al , 2020Riabovol et al 2019;Krasnytska et al 2022). Bioinformatics profiling identifies a glucose-related risk signature for the malignancy of glioma (Zhao et al 2017).…”

Comprehensive analysis based on glycolytic and glutaminolytic pathways signature for predicting prognosis and immunotherapy in ovarian cancer

“…Our study also found that INTS6 was a favorable prognosis gene for OV. SPAG4, was also defined as a novel potential cancer marker 55 , however, the level of SPAG4 expression was decreased in glioma cells treated by glutamine deprivation 56 . ZNF316 has been identified as a transcription factor, although its specific role in OV remains uncertain.…”

High MEIS3 Expression Indicates a Poor Prognosis for Patients with Stage II/III Colorectal Cancer

“…Hirschsprung's disease, a hereditary disorder with congenital megacolon as its primary symptom, is caused by MEIS3 deficiency and affects children and newborns [24]. Although there have been reports about the role of MEIS3 in hepatocellular carcinoma and glioma, whether and how MEIS3 participates in the progression of gut tumors is currently unknown [25,26].…”