PTEN/AKT signaling mediates chemoresistance in refractory acute myeloid leukemia through enhanced glycolysis

Ryu,Min   Jeong; Han,Jeongsu; Kim,Soo   Jeong; Lee,Min   Joung; Ju,Xianshu; Lee,Yu  Lim; Son,Jeong   Hwan; Cui,Jianchen; Jang,Yunseon; Chung,Woosuk; Song,Ik‑Chan; Kweon,Gi   Ryang; Heo,Jun   Young

doi:10.3892/or.2019.7308

PTEN/AKT signaling mediates chemoresistance in refractory acute myeloid leukemia through enhanced glycolysis

Authors:
- Min Jeong Ryu
- Jeongsu Han
- Soo Jeong Kim
- Min Joung Lee
- Xianshu Ju
- Yu Lim Lee
- Jeong Hwan Son
- Jianchen Cui
- Yunseon Jang
- Woosuk Chung
- Ik‑Chan Song
- Gi Ryang Kweon
- Jun Young Heo
View Affiliations

Affiliations: Department of Biochemistry, College of Medicine, Chungnam National University School of Medicine, Daejeon 35015, Republic of Korea, Department of Medical Science, Chungnam National University School of Medicine, Daejeon 35015, Republic of Korea, Brain Research Institute, Chungnam National University School of Medicine, Daejeon 35015, Republic of Korea, Department of Internal Medicine, Chungnam National University Hospital, Daejeon 35015, Republic of Korea, Department of Biochemistry, College of Medicine, Chungnam National University School of Medicine, Daejeon 35015, Republic of Korea
Published online on: September 12, 2019 https://doi.org/10.3892/or.2019.7308
Pages: 2149-2158

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Primary refractory acute myeloid leukemia (AML) and early recurrence of leukemic cells are among the most difficult hurdles to overcome in the treatment of AML. Moreover, uncertainties surrounding the molecular mechanism underlying refractory AML pose a challenge when it comes to developing novel therapeutic drugs. However, accumulating evidence suggests a contribution of phosphatase and tensin homolog (PTEN)/protein kinase B (AKT) signaling to the development of refractory AML. To assess PTEN/AKT signaling in AML, two types of AML cell lines were evaluated, namely control HL60 cells and KG1α cells, a refractory AML cell line that is resistant to idarubicin and cytarabine (AraC) treatment. Changes in the expression level of glycolysis‑ and mitochondrial oxidative phosphorylation‑related genes and proteins were evaluated by reverse transcription‑quantitative polymerase chain reaction and western blot analyses, respectively. The mitochondrial oxygen consumption and extracellular acidification rates were measured using an XF24 analyzer. CCK8 assay and Annexin V/PI staining were used to analyze cell viability and cellular apoptosis, respectively. The PTEN protein was found to be depleted, whereas AKT phosphorylation levels were elevated in KG1α cells compared with HL60 cells. These changes were associated with increased expression of glucose transporter 1 and hexokinase 2, and increased lactate production. AKT inhibition decreased the proliferation of KG1α cells and decreased extracellular acidification without affecting HL60 cells. Notably, AKT inhibition increased the susceptibility of KG1α cells to chemotherapy with idarubicin and AraC. Taken together, the findings of the present study indicate that activation of AKT by PTEN deficiency sustains the refractory AML status through enhancement of glycolysis and mitochondrial respiration, effects that may be rescued by inhibiting AKT activity.

View Figures

View References

1	Döhner H, Weisdorf DJ and Bloomfield CD: Acute Myeloid Leukemia. N Engl J Med. 373:1136–1152. 2015. View Article : Google Scholar : PubMed/NCBI
2	Gale RP: Advances in the treatment of acute myelogenous leukemia. N Engl J Med. 300:1189–1199. 1979. View Article : Google Scholar : PubMed/NCBI
3	Sakashita A, Hattori T, Miller CW, Suzushima H, Asou N, Takatsuki K and Koeffler HP: Mutations of the p53 gene in adult T-cell leukemia. Blood. 79:477–480. 1992.PubMed/NCBI
4	Kimby E, Nygren P and Glimelius B; SBU-group: Swedish Council of Technology Assessment in Health Care: A systematic overview of chemotherapy effects in acute myeloid leukaemia. Acta Oncol. 40:231–252. 2001. View Article : Google Scholar : PubMed/NCBI
5	Mohammadi M, Cao Y, Glimelius I, Bottai M, Eloranta S and Smedby KE: The impact of comorbid disease history on all-cause and cancer-specific mortality in myeloid leukemia and myeloma-a Swedish population-based study. BMC Cancer. 15:8502015. View Article : Google Scholar : PubMed/NCBI
6	De Kouchkovsky I and Abdul-Hay M: Acute myeloid leukemia: a comprehensive review and 2016 update'. Blood Cancer J. 6:e4412016. View Article : Google Scholar : PubMed/NCBI
7	Stone RM, Moser B, Sanford B, Schulman P, Kolitz JE, Allen S, Stock W, Galinsky I, Vij R, Marcucci G, Hurd D, et al: High dose cytarabine plus gemtuzumab ozogamicin for patients with relapsed or refractory acute myeloid leukemia: Cancer and leukemia Group B study 19902. Leuk Res. 35:329–333. 2011. View Article : Google Scholar : PubMed/NCBI
8	Aziz H, Ping CY, Alias H, Ab Mutalib NS and Jamal R: Gene mutations as emerging biomarkers and therapeutic targets for relapsed acute myeloid leukemia. Frontiers in Pharmacology. 8:8972017. View Article : Google Scholar : PubMed/NCBI
9	Levis M: Midostaurin approved for FLT3-mutated AML. Blood. 129:3403–3406. 2017. View Article : Google Scholar : PubMed/NCBI
10	Stein EM, DiNardo CD, Pollyea DA, Fathi AT, Roboz GJ, Altman JK, Stone RM, DeAngelo DJ, Levine RL, Flinn IW, et al: Enasidenib in mutant IDH2 relapsed or refractory acute myeloid leukemia. Blood. 130:722–731. 2017. View Article : Google Scholar : PubMed/NCBI
11	Lagadinou ED, Sach A, Callahan K, Rossi RM, Neering SJ, Minhajuddin M, Ashton JM, Pei S, Grose V, O'Dwyer KM, et al: BCL-2 inhibition targets oxidative phosphorylation and selectively eradicates quiescent human leukemia stem cells. Cell Stem Cell. 12:329–341. 2013. View Article : Google Scholar : PubMed/NCBI
12	Škrtić M, Sriskanthadevan S, Jhas B, Gebbia M, Wang X, Wang Z, Hurren R, Jitkova Y, Gronda M, Maclean N, et al: Inhibition of mitochondrial translation as a therapeutic strategy for human acute myeloid leukemia. Cancer Cell. 20:674–688. 2011. View Article : Google Scholar : PubMed/NCBI
13	Kuntz EM, Baquero P, Michie AM, Dunn K, Tardito S, Holyoake TL, Helgason GV and Gottlieb E: Targeting mitochondrial oxidative phosphorylation eradicates therapy-resistant chronic myeloid leukemia stem cells. Nat Med. 23:1234–1240. 2017. View Article : Google Scholar : PubMed/NCBI
14	Manning BD and Toker A: AKT/PKB Signaling: Navigating the network. Cell. 169:381–405. 2017. View Article : Google Scholar : PubMed/NCBI
15	Hawkins PT and Stephens LR: PI3K signalling in inflammation. Biochim Biophys Acta. 1851:882–897. 2015. View Article : Google Scholar : PubMed/NCBI
16	Zhao HF, Wang J and Tony To SS: The phosphatidylinositol 3-kinase/Akt and c-Jun N-terminal kinase signaling in cancer: Alliance or contradiction? (Review). Int J Oncol. 47:429–436. 2015. View Article : Google Scholar : PubMed/NCBI
17	Kang YH, Lee HS and Kim WH: Promoter methylation and silencing of PTEN in gastric carcinoma. Lab Invest. 82:285–291. 2002. View Article : Google Scholar : PubMed/NCBI
18	García JM, Silva J, Peña C, Garcia V, Rodríguez R, Cruz MA, Cantos B, Provencio M, España P and Bonilla F: Promoter methylation of the PTEN gene is a common molecular change in breast cancer. Genes Chromosomes Cancer. 41:117–124. 2004. View Article : Google Scholar : PubMed/NCBI
19	Alvarez-Nunez F, Bussaglia E, Mauricio D, Ybarra J, Vilar M, Lerma E, de Leiva A and Matias-Guiu X; Thyroid Neoplasia Study Group, : PTEN promoter methylation in sporadic thyroid carcinomas. Thyroid. 16:17–23. 2006. View Article : Google Scholar : PubMed/NCBI
20	Liu TC, Lin PM, Chang JG, Lee JP, Chen TP and Lin SF: Mutation analysis of PTEN/MMAC1 in acute myeloid leukemia. Am J Hematol. 63:170–175. 2000. View Article : Google Scholar : PubMed/NCBI
21	Huang X, Li D, Li T, Zhao BO and Chen X: Prognostic value of the expression of phosphatase and tensin homolog and CD44 in elderly patients with refractory acute myeloid leukemia. Oncol Lett. 10:103–110. 2015. View Article : Google Scholar : PubMed/NCBI
22	Yilmaz OH, Valdez R, Theisen BK, Guo W, Ferguson DO, Wu H and Morrison SJ: Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature. 441:475–482. 2006. View Article : Google Scholar : PubMed/NCBI
23	Zhang J, Grindley JC, Yin T, Jayasinghe S, He XC, Ross JT, Haug JS, Rupp D, Porter-Westpfahl KS, Wiedemann LM, et al: PTEN maintains haematopoietic stem cells and acts in lineage choice and leukaemia prevention. Nature. 441:518–522. 2006. View Article : Google Scholar : PubMed/NCBI
24	Fragoso R and Barata JT: PTEN and leukemia stem cells. Adv Biol Regul. 56:22–29. 2014. View Article : Google Scholar : PubMed/NCBI
25	Wang L, Xiong H, Wu F, Zhang Y, Wang J, Zhao L, Guo X, Chang LJ, Zhang Y, You MJ, et al: Hexokinase 2-mediated Warburg effect is required for PTEN- and p53-deficiency-driven prostate cancer growth. Cell Rep. 8:1461–1474. 2014. View Article : Google Scholar : PubMed/NCBI
26	Li Y, He L, Zeng N, Sahu D, Cadenas E, Shearn C, Li W and Stiles BL: Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) signaling regulates mitochondrial biogenesis and respiration via estrogen-related receptor α (ERRα). J Biol Chem. 288:25007–25024. 2013. View Article : Google Scholar : PubMed/NCBI
27	Pietkiewicz S, Schmidt JH and Lavrik IN: Quantification of apoptosis and necroptosis at the single cell level by a combination of Imaging Flow Cytometry with classical Annexin V/propidium iodide staining. J Immunol Methods. 423:99–103. 2015. View Article : Google Scholar : PubMed/NCBI
28	She M, Niu X, Chen X, Li J, Zhou M, He Y, Le Y and Guo K: Resistance of leukemic stem-like cells in AML cell line KG1a to natural killer cell-mediated cytotoxicity. Cancer Lett. 318:173–179. 2012. View Article : Google Scholar : PubMed/NCBI
29	Wiernik PH, Banks PL, Case DC Jr, Arlin ZA, Periman PO, Todd MB, Ritch PS, Enck RE and Weitberg AB: Cytarabine plus idarubicin or daunorubicin as induction and consolidation therapy for previously untreated adult patients with acute myeloid-leukemia. Blood. 79:313–319. 1992.PubMed/NCBI
30	Gallay N, Dos Santos C, Cuzin L, Bousquet M, Simmonet Gouy V, Chaussade C, Attal M, Payrastre B, Demur C and Récher C: The level of AKT phosphorylation on threonine 308 but not on serine 473 is associated with high-risk cytogenetics and predicts poor overall survival in acute myeloid leukaemia. Leukemia. 23:1029–1038. 2009. View Article : Google Scholar : PubMed/NCBI
31	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
32	Barthel A, Okino ST, Liao J, Nakatani K, Li J, Whitlock JP Jr and Roth RA: Regulation of GLUT1 gene transcription by the serine/threonine kinase Akt1. J Biol Chem. 274:20281–20286. 1999. View Article : Google Scholar : PubMed/NCBI
33	Xu DD, Wang Y, Zhou PJ, Qin SR, Zhang R, Zhang Y, Xue X, Wang J, Wang X, Chen HC, et al: The IGF2/IGF1R/nanog signaling pathway regulates the proliferation of acute myeloid leukemia stem cells. Front Pharmacol. 9:6872018. View Article : Google Scholar : PubMed/NCBI
34	Matkovic K, Brugnoli F, Bertagnolo V, Banfic H and Visnjic D: The role of the nuclear Akt activation and Akt inhibitors in all-trans-retinoic acid-differentiated HL-60 cells. Leukemia. 20:941–951. 2006. View Article : Google Scholar : PubMed/NCBI
35	Thol F, Schlenk RF, Heuser M and Ganser A: How I treat refractory and early relapsed acute myeloid leukemia. Blood. 126:319–327. 2015. View Article : Google Scholar : PubMed/NCBI
36	Bose P, Vachhani P and Cortes JE: Treatment of relapsed/refractory acute myeloid leukemia. Curr Treat Options Oncol. 18:172017. View Article : Google Scholar : PubMed/NCBI
37	Zhou Y, Tozzi F, Chen J, Fan F, Xia L, Wang J, Gao G, Zhang A, Xia X, Brasher H, et al: Intracellular ATP levels are a pivotal determinant of chemoresistance in colon cancer cells. Cancer Res. 72:304–314. 2012. View Article : Google Scholar : PubMed/NCBI
38	Song K, Li M, Xu X, Xuan LI, Huang G and Liu Q: Resistance to chemotherapy is associated with altered glucose metabolism in acute myeloid leukemia. Oncol Lett. 12:334–342. 2016. View Article : Google Scholar : PubMed/NCBI
39	Tanioka M, Sakai K, Sudo T, Sakuma T, Kajimoto K, Hirokaga K, Takao S, Negoro S, Minami H, Nakagawa K and Nishio K: Transcriptional CCND1 expression as a predictor of poor response to neoadjuvant chemotherapy with trastuzumab in HER2-positive/ER-positive breast cancer. Breast Cancer Res Treat. 147:513–525. 2014. View Article : Google Scholar : PubMed/NCBI
40	Jordan CT: The leukemic stem cell. Best Pract Res Clin Haematol. 20:13–18. 2007. View Article : Google Scholar : PubMed/NCBI
41	Vander Heiden MG, Cantley LC and Thompson CB: Understanding the Warburg effect: The metabolic requirements of cell proliferation. Science. 324:1029–1033. 2009. View Article : Google Scholar : PubMed/NCBI
42	Warburg O: On the origin of cancer cells. Science. 123:309–314. 1956. View Article : Google Scholar : PubMed/NCBI
43	Weinhouse S: On respiratory impairment in cancer cells. Science. 124:267–269. 1956. View Article : Google Scholar : PubMed/NCBI
44	Zu XL and Guppy M: Cancer metabolism: Facts, fantasy, and fiction. Biochem Biophys Res Commun. 313:459–465. 2004. View Article : Google Scholar : PubMed/NCBI
45	Li X, Jiang Y, Meisenhelder J, Yang W, Hawke DH, Zheng Y, Xia Y, Aldape K, He J, Hunter T, et al: Mitochondria-translocated PGK1 functions as a protein kinase to coordinate Glycolysis and the TCA Cycle in tumorigenesis. Mol Cell. 61:705–719. 2016. View Article : Google Scholar : PubMed/NCBI
46	Mates JM, Segura JA, Campos-Sandoval JA, Lobo C, Alonso L, Alonso FJ and Márquez J: Glutamine homeostasis and mitochondrial dynamics. Int J Biochem Cell Biol. 41:2051–2061. 2009. View Article : Google Scholar : PubMed/NCBI
47	Meng M, Chen S, Lao T, Liang D and Sang N: Nitrogen anabolism underlies the importance of glutaminolysis in proliferating cells. Cell Cycle. 9:3921–3932. 2010. View Article : Google Scholar : PubMed/NCBI
48	Fan J, Kamphorst JJ, Mathew R, Chung MK, White E, Shlomi T and Rabinowitz JD: Glutamine-driven oxidative phosphorylation is a major ATP source in transformed mammalian cells in both normoxia and hypoxia. Mol Syst Biol. 9:7122013. View Article : Google Scholar : PubMed/NCBI
49	Farge T, Saland E, de Toni F, Aroua N, Hosseini M, Perry R, Bosc C, Sugita M, Stuani L, Fraisse M, Scotland S, et al: Chemotherapy-resistant human acute myeloid leukemia cells are not enriched for leukemic stem cells but require oxidative metabolism. Cancer Discov. 7:716–735. 2017. View Article : Google Scholar : PubMed/NCBI
50	Gojo I, Perl A, Luger S, Baer MR, Norsworthy KJ, Bauer KS, Tidwell M, Fleckinger S, Carroll M and Sausville EA: Phase I study of UCN-01 and perifosine in patients with relapsed and refractory acute leukemias and high-risk myelodysplastic syndrome. Invest New Drugs. 31:1217–1227. 2013. View Article : Google Scholar : PubMed/NCBI
51	Sampath D, Malik A, Plunkett W, Nowak B, Williams B, Burton M, Verstovsek S, Faderl S, Garcia-Manero G, List AF, et al: Phase I clinical, pharmacokinetic, and pharmacodynamic study of the Akt-inhibitor triciribine phosphate monohydrate in patients with advanced hematologic malignancies. Leuk Res. 37:1461–1467. 2013. View Article : Google Scholar : PubMed/NCBI