Concurrent induction of apoptosis and necroptosis in apigenin‑treated malignant mesothelioma cells: Reversal of Warburg effect through Akt inhibition and p53 upregulation

Lee,Yoon-Jin; Park,Kwan-Sik; Heo,Su-Hak; Cho,Moon-Kyun; Lee,Sang-Han

doi:10.3892/or.2023.8548

Concurrent induction of apoptosis and necroptosis in apigenin‑treated malignant mesothelioma cells: Reversal of Warburg effect through Akt inhibition and p53 upregulation

Authors:
- Yoon-Jin Lee
- Kwan-Sik Park
- Su-Hak Heo
- Moon-Kyun Cho
- Sang-Han Lee
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Affiliations: Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheonan 31151, Republic of Korea, Department of Medicinal Bioscience, College of Biomedical and Health Science, Konguk University Glocal Campus, Chungju 27478, Republic of Korea, Department of Dermatology, Soonchunhyang University Seoul Hospital, Seoul 04401, Republic of Korea
Published online on: April 19, 2023 https://doi.org/10.3892/or.2023.8548
Article Number: 111
Copyright: © Lee et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A high dependence on aerobic glycolysis, known as the Warburg effect, is one of the metabolic features exhibited by tumor cells. Therefore, targeting glycolysis is becoming a very promising strategy for the development of anticancer drugs. In the present study, it was investigated whether pre‑adaptation of malignant mesothelioma (MM) cells to an acidic environment was associated with a metabolic shift to the Warburg phenotype in energy production, and whether apigenin targets acidosis‑driven metabolic reprogramming. Cell viability, glycolytic activity, Annexin V‑PE binding activity, reactive oxygen species (ROS) levels, mitochondrial membrane potential, ATP content, western blot analysis and spheroid viability were assessed in the present study. MM cells pre‑adapted to lactic acid were resistant to the anticancer drug gemcitabine, increased Akt activation, downregulated p53 expression, and upregulated rate‑limiting enzymes in glucose metabolism compared with their parental cells. Apigenin treatment increased cytotoxicity, Akt inactivation and p53 upregulation. Apigenin also reduced glucose uptake along with downregulation of key regulatory enzymes in glycolysis, increased ROS levels with loss of mitochondrial membrane potential, and downregulated the levels of complexes I, III and IV in the mitochondrial electron transport chain with intracellular ATP depletion, resulting in upregulation of molecules mediating apoptosis and necroptosis. Apigenin‑induced alterations of cellular responses were similar to those of Akt inactivation by Ly294002. Overall, the present results provide mechanistic evidence supporting the anti‑glycolytic and cytotoxic role of apigenin via inhibition of the PI3K/Akt signaling pathway and p53 upregulation.

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1	Warburg O: The metabolism of carcinoma cells. J Cancer Res. 9:148–163. 1925. View Article : Google Scholar
2	Tarrado-Castellarnau M, de Atauri P and Cascante M: Oncogenic regulation of tumor metabolic reprogramming. Oncotarget. 7:62726–62753. 2016. View Article : Google Scholar : PubMed/NCBI
3	Liberti MV and Locasale JW: The Warburg effect: How does it benefit cancer cells? Trends Biochem Sci. 41:211–218. 2016. View Article : Google Scholar : PubMed/NCBI
4	Eales KL, Hollinshead KER and Tennantm DA: Hypoxia and metabolic adaptation of cancer cells. Oncogenesis. 5:e1902016. View Article : Google Scholar : PubMed/NCBI
5	de la Cruz-López KG, Castro-Muñoz LJ, Reyes-Hernández DO, García-Carrancá A and Manzo-Merino J: Lactate in the regulation of tumor microenvironment and therapeutic approaches. Front Oncol. 9:11432019. View Article : Google Scholar : PubMed/NCBI
6	Engelman JA, Luo J and Cantley LC: The evolution of phosphatidylinositol 3-kinases as regulators of growth and metabolism. Nat Rev Genet. 7:606–619. 2006. View Article : Google Scholar : PubMed/NCBI
7	Courtnay R, Ngo DC, Malik N, Ververis K, Tortorella SM and Karagiannis TC: Cancer metabolism and the Warburg effect: The role of HIF-1 and PI3K. Mol Biol Rep. 42:841–851. 2015. View Article : Google Scholar : PubMed/NCBI
8	Wu Z, Wu J, Zhao Q, Fu S and Jin J: Emerging roles of aerobic glycolysis in breast cancer. Clin Transl Oncol. 22:631–646. 2020. View Article : Google Scholar : PubMed/NCBI
9	Elstrom RL, Bauer DE, Buzzai M, Karnauskas R, Harris MH, Plas DR, Zhuang H, Cinalli RM, Alavi A, Rudin CM and Thompson CB: Akt stimulates aerobic glycolysis in cancer cells. Cancer Res. 64:3892–3899. 2004. View Article : Google Scholar : PubMed/NCBI
10	Urso L, Cavallari I, Sharova E, Ciccarese F, Pasello G and Ciminale V: Metabolic rewiring and redox alterations in malignant pleural mesothelioma. Br J Cancer. 122:52–61. 2020. View Article : Google Scholar : PubMed/NCBI
11	Abraham AG and O'Neill E: PI3K/Akt-mediated regulation of p53 in cancer. Biochem Soc Trans. 42:798–803. 2014. View Article : Google Scholar : PubMed/NCBI
12	Naderali E, Valipour B, Khaki AA, Rad JS, Alihemmati A, Rahmati M and Charoudeh HN: Positive effects of PI3K/Akt signaling inhibition on PTEN and p53 in prevention of acute lymphoblastic leukemia tumor cells. Adv Pharm Bull. 9:470–480. 2019. View Article : Google Scholar : PubMed/NCBI
13	Song M, Bode AM, Dong Z and Lee MH: AKT as a therapeutic target for cancer. Cancer Res. 79:1019–1031. 2019. View Article : Google Scholar : PubMed/NCBI
14	Ahmed SA, Parama D, Daimari E, Girisa S, Banik K, Harsha C, Dutta U and Kunnumakkara AB: Rationalizing the therapeutic potential of apigenin against cancer. Life Sci. 267:1188142021. View Article : Google Scholar : PubMed/NCBI
15	Madunić J, Madunić IV, Gajski G, Popić J and Garaj-Vrhovac V: Apigenin: A dietary flavonoid with diverse anticancer properties. Cancer Lett. 413:11–22. 2018. View Article : Google Scholar : PubMed/NCBI
16	Tong X and Pelling JC: Targeting the PI3K/Akt/mTOR axis by apigenin for cancer prevention. Anticancer Agents Med Chem. 13:971–978. 2013. View Article : Google Scholar : PubMed/NCBI
17	Masuelli L, Benvenuto M, Mattera R, Di Stefano E, Zago E, Taffera G, Tresoldi I, Giganti MG, Frajese GV, Berardi G, et al: In vitro and in vivo anti-tumoral effects of the flavonoid apigenin in malignant mesothelioma. Front Pharmacol. 8:3732017. View Article : Google Scholar : PubMed/NCBI
18	Lee YJ, Park KS, Nam HS, Cho MK and Lee SH: Apigenin causes necroptosis by inducing ROS accumulation, mitochondrial dysfunction, and ATP depletion in malignant mesothelioma cells. Korean J Physiol Pharmacol. 24:493–502. 2020. View Article : Google Scholar : PubMed/NCBI
19	Xu RH, Pelicano H, Zhou Y, Carew JS, Feng L, Bhalla KN, Keating MJ and Huang P: Inhibition of glycolysis in cancer cells: A novel strategy to overcome drug-resistance associated with mitochondrial respiratory defect and hypoxia. Cancer Res. 65:613–621. 2005. View Article : Google Scholar : PubMed/NCBI
20	Geschwind JF, Ko YH, Torbenson MS, Magee C and Pedersen PL: Novel therapy for liver cancer: Direct intraarterial injection of a potent inhibitor of ATP production. Cancer Res. 62:3909–3913. 2002.PubMed/NCBI
21	Leist M, Single B, Castoldi AF, Kühnle S and Nicotera P: Intracellular adenosine triphosphate (ATP) concentration: A switch in the decision between apoptosis and necrosis. J Exp Med. 185:1481–1486. 1997. View Article : Google Scholar : PubMed/NCBI
22	Singhal S, Wiewrodt R, Malden LD, Amin KM, Matzie K, Friedberg J, Kucharczuk JC, Litzky LA, Johnson SW, Kaiser LR and Albelda SM: Gene expression profiling of malignant mesothelioma. Clin Cancer Res. 9:3080–3097. 2003.PubMed/NCBI
23	Bonelli M, Terenziani R, Zoppi S, Fumarola C, La Monica S, Cretella D, Alfieri R, Cavazzoni A, Digiacomo G, Galetti M and Petronini PG: Dual inhibition of CDK4/6 and PI3K/AKT/mTOR signaling impairs energy metabolism in MPM cancer cells. Int J Mol Sci. 21:51652020. View Article : Google Scholar : PubMed/NCBI
24	Lee YJ and Lee SH: Pro-oxidant activity of sulforaphane and cisplatin potentiates apoptosis and simultaneously promotes autophagy in malignant mesothelioma cells. Mol Med Rep. 16:2133–2141. 2017. View Article : Google Scholar : PubMed/NCBI
25	Kozlov AM, Lone A, Betts DH and Cumming RC: Lactate preconditioning promotes a HIF-1α-mediated metabolic shift from OXPHOS to glycolysis in normal human diploid fibroblasts. Sci Rep. 10:83882020. View Article : Google Scholar : PubMed/NCBI
26	Ma L and Zong X: Metabolic symbiosis in chemoresistance: Refocusing the role of aerobic glycolysis. Front Oncol. 10:52020. View Article : Google Scholar : PubMed/NCBI
27	He J, Xie G, Tong J, Peng Y, Huang H, Li J, Wang N and Liang H: Overexpression of microRNA-122 re-sensitizes 5-FU-resistant colon cancer cells to 5-FU through the inhibition of PKM2 in vitro and in vivo. Cell Biochem Biophys. 70:1343–1350. 2014. View Article : Google Scholar : PubMed/NCBI
28	Pérez-Tomás R and Pérez-Guillén I: Lactate in the tumor microenvironment: An essential molecule in cancer progression and treatment. Cancers (Basel). 12:32442020. View Article : Google Scholar : PubMed/NCBI
29	Ruan GX and Kazlauskas A: Lactate engages receptor tyrosine kinases Axl, Tie2, and vascular endothelial growth factor receptor 2 to activate phosphoinositide 3-kinase/Akt and promote angiogenesis. J Biol Chem. 288:21161–21172. 2013. View Article : Google Scholar : PubMed/NCBI
30	De Saedeleer CJ, Copetti T, Porporato PE, Verrax J, Feron O and Sonveaux P: Lactate activates HIF-1 in oxidative but not in Warburg-phenotype human tumor cells. PLoS One. 7:e465712012. View Article : Google Scholar : PubMed/NCBI
31	Fukumura D, Xu L, Chen Y, Gohongi T, Seed B and Jain RK: Hypoxia and acidosis independently up-regulate vascular endothelial growth factor transcription in brain tumors in vivo. Cancer Res. 61:6020–6024. 2001.PubMed/NCBI
32	Zhuo B, Li Y, Li Z, Qin H, Sun Q, Zhang F, Shen Y, Shi Y and Wang R: PI3K/Akt signaling mediated hexokinase-2 expression inhibits cell apoptosis and promotes tumor growth in pediatric osteosarcoma. Biochem Biophys Res Commun. 464:401–406. 2015. View Article : Google Scholar : PubMed/NCBI
33	Simabuco FM, Morale MG, Pavan ICB, Morelli AP, Silva FR and Tamura RE: p53 and metabolism: From mechanism to therapeutics. Oncotarget. 9:23780–23823. 2018. View Article : Google Scholar : PubMed/NCBI
34	Ma W, Sung HJ, Park JY, Matoba S and Hwang PM: A pivotal role for p53: Balancing aerobic respiration and glycolysis. J Bioenerg Biomembr. 39:243–246. 2007. View Article : Google Scholar : PubMed/NCBI
35	Xue C, Gu X, Li G, Bao Z and Li L: Mitochondrial mechanisms of necroptosis in liver diseases. Int J Mol Sci. 22:662020. View Article : Google Scholar : PubMed/NCBI
36	Edmondson R, Broglie JJ, Adcock AF and Yang L: Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay Drug Dev Technol. 12:207–218. 2014. View Article : Google Scholar : PubMed/NCBI
37	Chitcholtan K, Sykes P and Evans J: The resistance of intracellular mediators to doxorubicin and cisplatin are distinct in 3D and 2D endometrial cancer. J Transl Med. 10:382012. View Article : Google Scholar : PubMed/NCBI