bims-meluca 2025-06-15 papers

Biochim Biophys Acta Mol Basis Dis. 2025 Jun 05. pii: S0925-4439(25)00297-2. [Epub ahead of print] 167949

Metabolic profile of human non-small cell lung cancer cells through combined 13C and 2H NMR.

Ludgero C Tavares, Ricardo Amorim, José Teixeira, Paulo J Oliveira, Rui A Carvalho.

The metabolic remodeling occurring in carcinogenesis cells is firmly established. However, to understand the connection between the cellular metabolic profile and carcinogene sis, an accurate measurement of metabolic fluxes is required. In order to quantify the fluxes in these metabolic pathways, stable isotopes tracers and nuclear magnetic resonance (NMR) techniques were employed. For that purpose, two human non-small lung cancer cell lines (A549 and H1299) were used. For the quantification of carbon intermediary metabolism cells were grown in 13C labelled glucose while for de novo lipogenesis (DNL) assessment 2H2O was supplemented to the culture media. To better understand and characterize cellular bioenergetics, mitochondrial membrane potential, oxygen consumption, and energy charge were also assessed. Finally, to establish a bridge between metabolic fluxes and cancer proliferation, substrate dependency studies were performed. Several metabolic inhibitors were also tested, targeting glycolysis, TCA cycle, pentose phosphate pathway (PPP) and transaminases. Our results showed the occurrence of metabolic heterogeneity between the two non-small lung cancer cell lines: H1299 exhibited a relatively active TCA cycle, while A549 showed a more glycolytic phenotype. The overall mitochondrial bioenergetic parameters were in agreement with the metabolic profiles. The mitochondrial network was polarized and active in all cell lines, although the H1299 cell line exhibited higher basal oxygen consumption and spare respiratory capacity. Nonetheless, DNL rate did not differ in H1299 and A549 lung cancer cell lines. Additionally, α-ketoglutarate availability was proven a key determinant for H1299 non-small cell lung cancer cells survival and proliferation. In conclusion, this work revealed that cells derived from a lymph node metastasis (H1299) have a more active TCA cycle and altered oxidative stress levels when compared to cells derived from a primary tumor (A549). In the process, we successfully implemented a new 2H enrichment method for DNL assessment for the first time in in vitro cancer research.

Keywords: A549; Glycolysis; H1299; Krebs cycle; Lipogenesis; NADPH; Stable isotope tracers

DOI: https://doi.org/10.1016/j.bbadis.2025.167949

Int J Mol Sci. 2025 May 23. pii: 5014. [Epub ahead of print]26(11):

Modulation of the Antitumor Response to Metformin, Caffeine, and Sodium Dichloroacetate by the Hypoxic Microenvironment in Lung Cancer Cells.

Misael Osmar Garcia-Martin, Manuel Castillejos-Lopez, Heriberto Prado-Garcia, Susana Romero-Garcia, Juan Carlos Huerta-Cruz, José Alberto Choreño-Parra, Georgina Gonzalez-Avila, Luz A Colín-Godínez, Daniel Paz-Gomez, Ángeles Carlos-Reyes, Victor Ruiz, Yair Romero, Edgar Flores-Soto, Juan Rodríguez-Silverio, Roberto Lara-Lemus, Rafael Velázquez-Cruz, Citlaltepetl Salinas-Lara, Luz María Torres-Espíndola, Arnoldo Aquino Gálvez.

Metformin, caffeine, and dichloroacetate (DCA) have shown antitumor effects. The hypoxic tumor microenvironment can modulate drug response. We aimed to analyze the interaction of metformin with caffeine or DCA in lung cancer cells (HCC827) under normoxia and hypoxia conditions. Cell viability was evaluated using the crystal violet assay after individual and combined drug treatment under normoxia (21% O2) and hypoxia (1% O2) conditions. Combination effects were analyzed using isobolographic analysis. The results show that under normoxia conditions, the combination of metformin with DCA (γ = 0.98 ± 0.35, p > 0.05) or caffeine (γ = 0.90 ± 0.34, p > 0.05) revealed additivity. However, in hypoxia, both combinations exhibited significant antagonism, with γ values appearing greater than one for metformin + DCA (γ = 4.20 ± 1.44, p < 0.05) and metformin + caffeine (γ = 2.88 ± 0.90, p < 0.05). Hypoxia significantly alters the pharmacological interaction of metformin with caffeine or DCA, which could limit their combined therapeutic potential in hypoxic tumors despite metformin's activity in this environment. The importance of considering tumor oxygenation status in the design of combined therapies for lung cancer is emphasized.

Keywords: DCA; HCC827; caffeine; hypoxia; isobolographic analysis; lung cancer; metformin

DOI: https://doi.org/10.3390/ijms26115014

Cell Oncol (Dordr). 2025 Jun 11.

GMPS inhibits the proliferation and migration of non-small cell lung cancer via the regulation of the DNMT 1/SERPINB2 axis.

Tingting Guo, Lei Liu, Lingyan Zeng, Ying Yang, Tingting Song, Huachang Zhao, Zhixin Qiu.

BACKGROUND: Non-small cell lung cancer (NSCLC) mainly includes lung squamous cell carcinoma and lung adenocarcinoma, and its extremely high morbidity and mortality are the main causes of poor prognosis in NSCLC patients. Therefore, it is particularly important to study the mechanisms associated with tumor proliferation and metastasis and explore new molecular targets of NSCLC. Studies have shown that Guanosine monophosphate synthase (GMPS) may serve as a potential drug target, but its biological function and molecular mechanism in NSCLC are still unknown. Therefore, it is urgently needed to investigate the molecular mechanisms of GMPS.
METHODS: We first analyzed 30 cases of lung adenocarcinoma, lung squamous carcinoma and adjacent tissues; Then, lentiviral technology was used to construct overexpressed or knocked out cell lines to verify the function of GMPS. Then, RNA sequencing and Western blot experiments were carried out in animal experiments to explore the mechanism of GMPS. Our experimental results suggest that GMPS plays an important role in the progression of NSCLC.
RESULTS: We found that GMPS was highly expressed in lung adenocarcinoma and lung squamous cell carcinoma tissues, and was associated with poor prognosis of patients. Down-regulation of GMPS inhibits tumor progression. And GMPS promotes lung cancer cell migration through the SERPINB2-uPA axis, and DNMT1 is an intermediate factor in GMPS regulating SERPINB2 expression. Our experimental results show that GMPS expression is associated with lung cancer invasion and migration.
CONCLUSIONS: Our findings revealed the correlation between GMPS and the prognosis of NSCLC at the tissue level. Secondly, GMPS can promote the progression of NSCLC. The molecular mechanism of GMPS affecting the metastasis of lung cancer cells was elucidated. These findings highlight the important role of GMPS in NSCLC, so as to provide new insights for the identification of new targets and lay a theoretical foundation for the clinical application of GMPS.

Keywords: DNMT1; GMPS; Migration; Non-small cell lung cancer; SERPINB2

DOI: https://doi.org/10.1007/s13402-025-01078-1