Hypoxia Changes Energy Metabolism and Growth Rate in Non-Small Cell Lung Cancer Cells

Abstract

Hypoxia occurs in 80% of non-small cell lung carcinoma (NSCLC) cases, leading to treatment resistance. Hypoxia’s effects on NSCLC energetics are not well-characterized. We evaluated changes in glucose uptake and lactate production in two NSCLC cell lines under hypoxia in conjunction with growth rate and cell cycle phase distribution. The cell lines A549 (p53 wt) and H358 (p53 null) were incubated under hypoxia (0.1% and 1% O2) or normoxia (20% O2). Glucose and lactate concentrations in supernatants were measured using luminescence assays. Growth kinetics were followed over seven days. Cell nuclei were stained with DAPI and nuclear DNA content was determined by flow cytometry to determine cell cycle phase. Gene expression under hypoxia was determined by RNA sequencing. Glucose uptake and lactate production under hypoxia were greater than under normoxia. They were also significantly greater in A549 compared to H358 cells. Faster energy metabolism in A549 cells was associated with a higher growth rate in comparison to H358 cells under both normoxia and hypoxia. In both cell lines, hypoxia significantly slowed down the growth rate compared to proliferation under normoxic conditions. Hypoxia led to redistribution of cells in the different cycle phases: cells in G1 increased and the G2 population decreased. Glucose uptake and lactate production increase under hypoxia in NSCLC cells indicated greater shunting of glucose into glycolysis rather than into oxidative phosphorylation compared to normoxia, making adenosine triphosphate (ATP) production less efficient. This may explain the redistribution of hypoxic cells in the G1 cell cycle phase and the time increase for cell doubling. Energy metabolism changes were more prominent in faster-growing A549 cells compared to slower-growing H358 cells, indicating possible roles for the p53 status and inherent growth rate of different cancer cells. In both cell lines, genes associated with cell motility, locomotion and migration were upregulated under chronic hypoxia, indicating a strong stimulus to escape hypoxic conditions.

Hypoxia: Intriguing Feature in Cancer Cell Biology

“…Copyright 2021, Frontiers Media SA, Angiogenesis Reproduced with permission [33] . Copyright 2022, Multidisciplinary Digital Publishing Institute (MDPI), Metabolism Reproduced with permission [37] . Copyright 2023, Multidisciplinary Digital Publishing Institute (MDPI), Immunoresponse Reproduced with permission [42] .…”

“…[33] Copyright 2022, Multidisciplinary Digital Publishing Institute (MDPI), Metabolism Reproduced with permission. [37] Copyright 2023, Multidisciplinary Digital Publishing Institute (MDPI), Immunoresponse Reproduced with permission. [42] Copyright 2016, Frontiers Media SA, Cell Death Reproduced with permission.…”

“…These findings highlight that hypoxia elevates glycolysis, diminishes mitochondrial respiration, and decreases ROS production in tumor cells (Figure 2). [36,37]…”

USP7 promotes non‐small‐cell lung cancer cell glycolysis and survival by stabilizing and activating c‐Abl

“… 37 Tumour cells are predisposed to produce ATP through glycolysis at both hypoxia and normoxia to promote tumour cell growth. 38 Hexokinases are the rate‐limiting enzymes in promoting glycolysism, and it has been upregulated in cancers by many factors such as activated ERBB2, RAS and AKT, as well as loss of p53. 39 HK2, a key member of the HK family, catalyzes the first committed step in glucose metabolism by phosphorylating glucose, and it is specifically expressed in many foetal tissues and cancer cells.…”

Differential Inhibition of Anaplerotic Pyruvate Carboxylation and Glutaminolysis-Fueled Anabolism Underlies Distinct Toxicity of Selenium Agents in Human Lung Cancer

“…In particular, the ability to block both PC and glutaminase by MSeA and selenite, but not by SeM, correlated well with their ability to inhibit cell proliferation and to induce ROS production. These effects of MSeA and selenite on anaplerosis led to reduced Krebs cycle activity, which in turn attenuated de novo synthesis of nucleotides and lipids, all required for supporting cell proliferation [ 93 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 ]. We have previously shown that PC is activated in vivo in NSCLC patient tumors [ 61 ], and that both PC and glutaminolysis are important for supporting lung cancer cell proliferation [ 91 ] .…”