Alpha ketoglutarate exerts a pro-osteogenic effect in osteoblast cell lines through activation of JNK and mTOR/S6K1/S6 signaling pathways

Żurek, Aleksandra; Mizerska-Kowalska, Magdalena; Sławińska-Brych, Adrianna; Kaławaj, Katarzyna; Bojarska‐Junak, Agnieszka; Kandefer‐Szerszeń, Martyna; Zdzisińska, Barbara

doi:10.1016/j.taap.2019.04.024

Cited by 23 publications

(24 citation statements)

References 52 publications

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“…Evidence from recent studies suggests that exogenous supplementation of AKG may exert anti-cancer effects against colorectal carcinoma or breast cancer [19,22]. Moreover, our recent study has revealed that exogenous AKG can inhibit cell proliferation and stimulate differentiation of normal osteoblasts [36]. However, the influence of AKG on osteosarcoma cell lines has not been studied so far.…”

Section: Discussionmentioning

confidence: 97%

“…The quantification of the intracellular levels of total and phosphorylated JNK, ERK1/2, p38, and AKT kinases and the contents of cyclin D1 and p21 proteins in the treated cells was carried out using the PathScan ® ELISA kits: Total SAPK/JNK Sandwich ELISA Kit, Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA Kit, Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit, Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit, Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit, Total Cyclin D1 Sandwich ELISA Kit, Total p21 Waf1/Cip1 Sandwich ELISA Kit (Cell Signaling Technology Danvers, MA, USA), and p38 MAPK alpha ELISA Kit (Abcam, Cambridge, UK) according to the manufacturer's instructions as described previously [36]. Briefly, the Saos-2 cells (1 × 10 6 cells/mL) were incubated in culture medium (2% FBS) without or with the selected concentrations of AKG (10,25, and 50 mM) in 10-cm diameter plastic plates.…”

Section: Pathscan Elisa Assaysmentioning

confidence: 99%

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Alpha Ketoglutarate Exerts In Vitro Anti-Osteosarcoma Effects through Inhibition of Cell Proliferation, Induction of Apoptosis via the JNK and Caspase 9-Dependent Mechanism, and Suppression of TGF-β and VEGF Production and Metastatic Potential of Cells

Kaławaj

Sławińska-Brych

Mizerska-Kowalska

et al. 2020

IJMS

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Osteosarcoma (OS) is the most common type of primary bone tumor. Currently, there are limited treatment options for metastatic OS. Alpha-ketoglutarate (AKG), i.e., a multifunctional intermediate of the Krebs cycle, is one of the central metabolic regulators of tumor fate and plays an important role in cancerogenesis and tumor progression. There is growing evidence suggesting that AKG may represent a novel adjuvant therapeutic opportunity in anti-cancer therapy. The present study was intended to check whether supplementation of Saos-2 and HOS osteosarcoma cell lines (harboring a TP53 mutation) with exogenous AKG exerted an anti-cancer effect. The results revealed that AKG inhibited the proliferation of both OS cell lines in a concentration-dependent manner. As evidenced by flow cytometry, AKG blocked cell cycle progression at the G1 stage in both cell lines, which was accompanied by a decreased level of cyclin D1 in HOS and increased expression of p21Waf1/Cip1 protein in Saos-2 cells (evaluated with the ELISA method). Moreover, AKG induced apoptotic cell death and caspase-3 activation in both OS cell lines (determined by cytometric analysis). Both the immunoblotting and cytometric analysis revealed that the AKG-induced apoptosis proceeded predominantly through activation of an intrinsic caspase 9-dependent apoptotic pathway and an increased Bax/Bcl-2 ratio. The apoptotic process in the AKG-treated cells was mediated via c-Jun N-terminal protein kinase (JNK) activation, as the specific inhibitor of this kinase partially rescued the cells from apoptotic death. In addition, the AKG treatment led to reduced activation of extracellular signal-regulated kinase (ERK1/2) and significant inhibition of cell migration and invasion in vitro concomitantly with decreased production of pro-metastatic transforming growth factor β (TGF-β) and pro-angiogenic vascular endothelial growth factor (VEGF) in both OS cell lines suggesting the anti-metastatic potential of this compound. In conclusion, we showed the anti-osteosarcoma potential of AKG and provided a rationale for a further study of the possible application of AKG in OS therapy.

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Section: Discussionmentioning

confidence: 97%

Section: Pathscan Elisa Assaysmentioning

confidence: 99%

Alpha Ketoglutarate Exerts In Vitro Anti-Osteosarcoma Effects through Inhibition of Cell Proliferation, Induction of Apoptosis via the JNK and Caspase 9-Dependent Mechanism, and Suppression of TGF-β and VEGF Production and Metastatic Potential of Cells

Kaławaj

Sławińska-Brych

Mizerska-Kowalska

et al. 2020

IJMS

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“…A recent study has also documented the antiaging effects of AKG in Drosophila, that also mediates its effects via mTOR inhibition [37]. However, AKG may activate mTOR signaling pathway [38,39]. In intestinal porcine epithelial cells, AKG activated mTOR to stimulate protein synthesis [40].…”

Section: Agingmentioning

confidence: 99%

“…There was a significant decrease in the C-terminal crosslinking telopeptide of type-I collagen (a biomarker of bone reabsorption) in patients receiving Ca-AKG supplement compared to those only receiving calcium [31]. The mechanism may be related to the effect of AKG on epigenetic regulation [39,53]. AKG increased bone mass, attenuated age-related mass bone loss, and stimulated osteogenic differentiation in 18-month-old mice via regulating histone methylation [53].…”

Section: Osteoporosismentioning

confidence: 99%

Alpha-Ketoglutarate dietary supplementation to improve health in humans

Gyanwali¹,

Lim²,

Soh³

et al. 2022

Trends in Endocrinology & Metabolism

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“…The degree of the extracellular matrix mineralization was determined using Alizarin Red S (ARS, Sigma) staining of cultured osteoblasts to detect bone nodules (calcium precipitates) as previously described [63]. The ARS-stained cell cultures were examined under a light microscope.…”

Section: Discussionmentioning

confidence: 99%

Betulin Promotes Differentiation of Human Osteoblasts In Vitro and Exerts an Osteoinductive Effect on the hFOB 1.19 Cell Line Through Activation of JNK, ERK1/2, and mTOR Kinases

et al. 2019

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Although betulin (BET), a naturally occurring pentacyclic triterpene, has a variety of biological activities, its osteogenic potential has not been investigated so far. The aim of this study was to assess the effect of BET on differentiation of human osteoblasts (hFOB 1.19 and Saos-2 cells) in vitro in osteogenic (with ascorbic acid as an osteogenic supplement) and osteoinductive (without an additional osteogenic supplement) conditions. Osteoblast differentiation was evaluated based on the mRNA expression (RT-qPCR) of Runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), type I collagen-α1 (COL1A1), and osteopontin (OPN). Additionally, ALP activity and production of COL1A1 (western blot analysis) and OPN (ELISA) were evaluated. The level of mineralization (calcium accumulation) was determined with Alizarin red S staining. BET upregulated the mRNA level of RUNX2 and the expression of other osteoblast differentiation markers in both cell lines (except the influence of BET on ALP expression/activity in the Saos-2 cells). Moreover, it increased mineralization in both cell lines in the osteogenic conditions. BET also increased the mRNA level of osteoblast differentiation markers in both cell lines (except for ALP in the Saos-2 cells) in the osteoinductive conditions, which was accompanied with increased matrix mineralization. The osteoinductive activity of BET in the hFOB 1.19 cells was probably mediated via activation of MAPKs (JNK and ERK1/2) and mTOR, as the specific inhibitors of these kinases abolished the BET-induced osteoblast differentiation. Our results suggest that BET has the potential to enhance osteogenesis.

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Alpha ketoglutarate exerts a pro-osteogenic effect in osteoblast cell lines through activation of JNK and mTOR/S6K1/S6 signaling pathways

Cited by 23 publications

References 52 publications

Alpha Ketoglutarate Exerts In Vitro Anti-Osteosarcoma Effects through Inhibition of Cell Proliferation, Induction of Apoptosis via the JNK and Caspase 9-Dependent Mechanism, and Suppression of TGF-β and VEGF Production and Metastatic Potential of Cells

Alpha Ketoglutarate Exerts In Vitro Anti-Osteosarcoma Effects through Inhibition of Cell Proliferation, Induction of Apoptosis via the JNK and Caspase 9-Dependent Mechanism, and Suppression of TGF-β and VEGF Production and Metastatic Potential of Cells

Alpha-Ketoglutarate dietary supplementation to improve health in humans

Betulin Promotes Differentiation of Human Osteoblasts In Vitro and Exerts an Osteoinductive Effect on the hFOB 1.19 Cell Line Through Activation of JNK, ERK1/2, and mTOR Kinases

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