Aldolase A promotes proliferation and G₁/S transition via the EGFR/MAPK pathway in non‐small cell lung cancer

Abstract

BackgroundOur previous study demonstrated that aldolase A (ALDOA) is overexpressed in clinical human lung squamous cell carcinoma and that ALDOA promotes epithelial–mesenchymal transition and tumorigenesis. The present study aimed to explore the function of ALDOA in the modulation of non-small cell lung cancer (NSCLC) proliferation and cell cycle progression and the potential mechanism.MethodsALDOA was knocked down by short hairpin RNA in H520 and H1299 cells. ALDOA was overexpressed with vectors carrying the full-length ALDOA sequence in H1299 and H157 cells. The proliferation capacities were assessed with immunohistochemical staining, Cell Counting Kit-8 and colony formation assays. The cell cycle distribution was examined by flow cytometry, and molecular alterations were determined by western blotting. Cell synchronization was induced with nocodazole. The stability of cyclin D1 mRNA was tested. The pyruvate kinase M2 and ALDOA protein distributions were examined. Aerobic glycolysis was evaluated with Cell Titer-Glo assay, glucose colorimetric assay and lactate colorimetric assay.ResultsALDOA knockdown inhibited the proliferation and G1/S transition in H520 cells. Conversely, ALDOA overexpression promoted the proliferation and G1/S transition in H157 cells. The cell cycle synchronization assay showed that ALDOA expression increased in the G1 phase and G1/S transition. Furthermore, ALDOA knockdown reduced cyclin D1 expression by regulating epidermal growth factor receptor/mitogen-activated protein kinase (EGFR/MAPK) pathway. Similar results were found in H1299 and H157 cells. The inhibition of mitogen-activated protein kinase kinase 1/2 prompted the nuclear distribution of ALDOA. Additionally, ALDOA knockdown reduced nuclear distribution of PKM2, the extracellular lactate and intracellular adenosine triphosphate concentrations and elevated the extracellular glucose concentration.ConclusionsALDOA contributed to activation of the EGFR/MAPK pathway, thus promoting cyclin D1 expression and enhancing proliferation and G1/S transition in NSCLC. Additionally, ALDOA facilitated NSCLC aerobic glycolysis.

Expression of genes in the16p11.2locus during human fetal cortical neurogenesis

“…ALDOA is much more abundant in the cytoplasm and we also found no clear relationship between cell body ALDOA levels and cell proliferation status. Nuclear ALDOA has been linked to cell proliferation (Mamczur et al 2010(Mamczur et al , 2013Fu et al 2018). While we were able to detect nuclear ALDOA we found no clear relationship between ALDOA protein levels and cell proliferation status.…”

“…The metabolic role of cytoplasmic ALDOA is well established, and ALDOA also has other non-glycolytic "moonlighting" roles such as regulating mitochondrial function and cyctoskeleton stability (Orosz et al 1988;Pagliaro and Taylor 1992;Kao et al 1999;Jewett and Sibley 2003;Buscaglia et al 2006). In addition to its cytoplasmic roles, ALDOA has been observed in the nucleus (Mamczur and Dzugaj 2008;Mamczur et al 2010Mamczur et al , 2013 where it has been suggested to impact cell cycle by positively regulating cyclin D1 expression to mediate 30 G1/S progression (Ritterson Lew and Tolan 2012;Fu et al 2018). Cell-culture studies show ALDOA sub-cellular localisation depends on the availability of energetic substrates, with addition of glucose driving ALDOA protein to the cytoplasm (Mamczur et al 2013).…”

“…Therefore, it is likely the primary role for ALDOA is metabolic when cells require, and have available to them, large amounts of energy. The majority of ALDOA studies have used highly abnormal cancer tissue, or artificial cell culture systems in which glycolytic enzymes have been shown to be increased (Ritterson Lew and Tolan 2012;Mamczur et al 2013;Fu et al 2018;Pollen et al 2019). How these observations of ALDOA in a variety of systems relate to its role in human cerebral cortex development is unclear.…”

Enrichment of Aldolase C Correlates with Low Non-Mutated IDH1 Expression and Predicts a Favorable Prognosis in Glioblastomas

“…Aldolase converts fructose 1-6, bisphosphate to glyceraldehyde 3-phosphate (G3P) and Dihydroxyacetone phosphate (DHAP), so they control fatty acid synthesis and glycolysis to further affect the Warburg effect or oxidative phosphorylation (OXPHOs). Otherwise, overexpression of ALDOA has been proven to be associated with poor overall outcome in colorectal and prostatic cancers, as well as lung non-small cell carcinoma [8,9,10]. ALDOA has a hypoxia response element (HRE) in the promoter region.…”

Antiproliferative and Antimetastatic Effects of Praeruptorin C on Human Non–Small Cell Lung Cancer through Inactivating ERK/CTSD Signalling Pathways

“…MAPK inhibitors, particularly JNK, enhance the cell death effects in H 2 O 2 -treated lung cancer cells by increasing superoxide anion and glutathione depletion [35]. The EGFR/MAPK pathway mediates aldolase A-promoted cyclin D1 expression and proliferation and G1/S transition in NSCLC [36]. Activation of the MAPK pathway by the long noncoding RNA TUC338 promotes the invasion of lung cancer [37].…”