ALDOC- and ENO2- driven glucose metabolism sustains 3D tumor spheroids growth regardless of nutrient environmental conditions: a multi-omics analysis

Abstract

Background Metastases are the major cause of cancer-related morbidity and mortality. By the time cancer cells detach from their primary site to eventually spread to distant sites, they need to acquire the ability to survive in non-adherent conditions and to proliferate within a new microenvironment in spite of stressing conditions that may severely constrain the metastatic process. In this study, we gained insight into the molecular mechanisms allowing cancer cells to survive and proliferate in an anchorage-independent manner, regardless of both tumor-intrinsic variables and nutrient culture conditions. Methods 3D spheroids derived from lung adenocarcinoma (LUAD) and breast cancer cells were cultured in either nutrient-rich or -restricted culture conditions. A multi-omics approach, including transcriptomics, proteomics, and metabolomics, was used to explore the molecular changes underlying the transition from 2D to 3D cultures. Small interfering RNA-mediated loss of function assays were used to validate the role of the identified differentially expressed genes and proteins in H460 and HCC827 LUAD as well as in MCF7 and T47D breast cancer cell lines. Results We found that the transition from 2D to 3D cultures of H460 and MCF7 cells is associated with significant changes in the expression of genes and proteins involved in metabolic reprogramming. In particular, we observed that 3D tumor spheroid growth implies the overexpression of ALDOC and ENO2 glycolytic enzymes concomitant with the enhanced consumption of glucose and fructose and the enhanced production of lactate. Transfection with siRNA against both ALDOC and ENO2 determined a significant reduction in lactate production and cell viability. Furthermore, both the number and size of spheroids produced by H460, HCC827, MCF7, and T47D cell lines were significantly reduced upon ALDOC and ENO2 knockdown. Conclusions Our results show that anchorage-independent survival and growth of cancer cells are supported by changes in genes and proteins that drive glucose metabolism towards an enhanced lactate production. Notably, this finding is valid regardless of the tumor type and nutrient environmental availability, thus suggesting the possible general involvement of this mechanism in cancer metastasis. The pan-cancer validation of this vulnerability could potentially help to slow or prevent cancer progression.

Biomimetic Nanomedicine Targeting Orchestrated Metabolism Coupled with Regulatory Factors to Disrupt the Metabolic Plasticity of Breast Cancer

“…Ttll7 encodes tubulin tyrosine ligase which affects the post-translational modification of tubulin and glutamylation. Aldoc encodes glycolytic enzymes that catalyze the reversible cleavage of fructose-1, 6-diphosphate, and fructose 1-phosphate to dihydroxyacetone phosphate and glyceraldehyde-3-phosphate or glyceraldehyde, respectively . Therefore, the transcription of glutamine- and glucose-related genes was significantly inhibited in breast cancer cells after treatment with RGFM.…”

“…Aldoc encodes glycolytic enzymes that catalyze the reversible cleavage of fructose-1, 6-diphosphate, and fructose 1-phosphate to dihydroxyacetone phosphate and glyceraldehyde-3-phosphate or glyceraldehyde, respectively. 26 Therefore, the transcription of glutamine-and glucose-related genes was significantly inhibited in breast cancer cells after treatment with RGFM. Krt13, id2, hspa4l, sfn, adamts8, ppan, and ccn2 are closely related to tumor proliferation, apoptosis, invasion and metastasis, which may be the corresponding response to metabolic dysfunction.…”

THER: Integrative Web Tool for Tumor Hypoxia Exploration and Research

“…Moreover, ENO2 was found to be significantly upregulated in six of the seven MCF7 cell line datasets (Figure 7C, Figure S1), which further validated the reliability of ENO2 as a candidate marker gene. ENO2 has been reported to function in hypoxia-associated tumors through the glycolytic pathway(33,38). Similarly, enrichment analysis showed that multiple glycolytic pathways were differential pathways in MCF7 cells (Figure 7E), and all of these pathways were involved in ENO2 (Figure 7F).…”

“…ENO2 encodes a glycolytic enzyme involved in the energy release phase of glycolysis(39,40). Under hypoxic conditions, breast cancer cells exhibit an enhanced glycolytic phenotype, resulting in upregulation of the expression of genes encoding glycolytic enzymes, including ENO2, and increased glycolytic energy production(38). Moreover, silencing ENO2 shows synergistic effects with anti-glycolytic drugs(41).…”