Effects of endoplasmic reticulum stress on autophagy and apoptosis of human leukemia cells via inhibition of the PI3K/AKT/mTOR signaling pathway

Li,Li‑Juan; Chai,Ye; Guo,Xiao‑Jia; Chu,Song‑Lin; Zhang,Lian‑Sheng

doi:10.3892/mmr.2018.8840

Effects of endoplasmic reticulum stress on autophagy and apoptosis of human leukemia cells via inhibition of the PI3K/AKT/mTOR signaling pathway

Authors:
- Li‑Juan Li
- Ye Chai
- Xiao‑Jia Guo
- Song‑Lin Chu
- Lian‑Sheng Zhang
View Affiliations

Affiliations: Department of Hematology, Lanzhou University Second Hospital, Lanzhou, Gansu 730000, P.R. China
Published online on: April 3, 2018 https://doi.org/10.3892/mmr.2018.8840
Pages: 7886-7892

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

The present study aimed to explore the regulatory effects of endoplasmic reticulum stress (ERS) on the phosphoinositide 3‑kinase (PI3K)/AKT serine/threonine kinase 1 (AKT)/mammalian target of rapamycin (mTOR) signaling pathway, and its subsequent effects on autophagy and apoptosis of human leukemia cells. Human leukemia cells were cultured and treated with various concentrations of tunicamycin for 0, 24, 48, 72 and 90 h. Subsequently, human leukemia cells were assigned into the ER activation group, which was treated with 100 ng/ml tunicamycin, the ER activation + TO901317 (PI3K inhibitor) group, and the control group. An MTT assay was conducted to detect cell proliferation. In addition, a monodansylcadaverine (MDC) assay was used to detect the formation of autophagosomes and Annexin V‑fluorescein isothiocyanate/propidium iodide double staining was used to examine cell apoptosis. Western blotting was performed to detect the expression levels of 78‑kDa glucose‑regulated protein (GRP78), phosphorylated (p)‑protein kinase R‑like endoplasmic reticulum kinase (PERK), p‑α subunit of eukaryotic initiation factor 2 (eIF2α), microtubule‑associated protein 1A/1B‑light chain 3 (LC3), caspase‑3, CCAAT‑enhancer‑binding protein homologous protein (CHOP), PI3K, AKT and mTOR. Treatment with 100 ng/ml tunicamycin for 72 h was considered the optimal condition for further experiments. Compared with in cells prior to treatment, human leukemia cells treated with tunicamycin exhibited increased expression of p‑PERK, p‑eIF2α and GRP78 after 72 h (P<0.05). In addition, the expression levels of mTOR, AKT and PI3K were decreased in the ER activation group compared with in the control and ER activation + TO901317 groups (P<0.05). Compared with in the control group, cell proliferation was inhibited and MDC fluorescence intensity was increased in the ER activation group (P<0.05). Furthermore, compared with in the control and ER activation + TO901317 groups, western blotting indicated that the expression levels of LC3‑II were increased in the ER activation group (P<0.05). The apoptotic rate was also higher in the ER activation group compared with in the control group (P<0.05), and caspase‑3 and CHOP expression was elevated in the ER activation group (P<0.05). These findings indicated that ERS may induce autophagy and apoptosis of human leukemia cells via inhibiting the PI3K/AKT/mTOR signaling pathway.

View Figures

View References

1	Xie Y, Davies SM, Xiang Y, Robison LL and Ross JA: Trends in leukemia incidence and survival in the United States (1973–1998). Cancer. 97:2229–2235. 2003. View Article : Google Scholar : PubMed/NCBI
2	Tower RL and Spector LG: The epidemiology of childhood leukemia with a focus on birth weight and diet. Crit Rev Clin Lab Sci. 44:203–242. 2007. View Article : Google Scholar : PubMed/NCBI
3	Ali NA, O'Brien JM Jr, Blum W, Byrd JC, Klisovic RB, Marcucci G, Phillips G, Marsh CB, Lemeshow S and Grever MR: Hyperglycemia in patients with acute myeloid leukemia is associated with increased hospital mortality. Cancer. 110:96–102. 2007. View Article : Google Scholar : PubMed/NCBI
4	Rajabli N, Naeimi-Tabeie M, Jahangirrad A, Sedaghat SM, Semnani S and Roshandel G: Epidemiology of leukemia and multiple myeloma in Golestan, Iran. Asian Pac J Cancer Prev. 14:2333–2336. 2013. View Article : Google Scholar : PubMed/NCBI
5	Rhomberg LR, Bailey LA, Goodman JE, Hamade AK and Mayfield D: Is exposure to formaldehyde in air causally associated with leukemia?-A hypothesis-based weight-of-evidence analysis. Crit Rev Toxicol. 41:555–621. 2011. View Article : Google Scholar : PubMed/NCBI
6	Yokoyama T, Miyazawa K, Naito M, Toyotake J, Tauchi T, Itoh M, Yuo A, Hayashi Y, Georgescu MM, Kondo Y, et al: Vitamin K2 induces autophagy and apoptosis simultaneously in leukemia cells. Autophagy. 4:629–640. 2008. View Article : Google Scholar : PubMed/NCBI
7	Ge J, Liu Y, Li Q, Guo X, Gu L, Ma ZG and Zhu YP: Resveratrol induces apoptosis and autophagy in T-cell acute lymphoblastic leukemia cells by inhibiting Akt/mTOR and activating p38-MAPK. Biomed Environ Sci. 26:902–911. 2013.PubMed/NCBI
8	Bao T, Smith BD and Karp JE: New agents in the treatment of acute myeloid leukemia: A snapshot of signal transduction modulation. Clin Adv Hematol Oncol. 3(287–296): 3022005.
9	LoPiccolo J, Blumenthal GM, Bernstein WB and Dennis PA: Targeting the PI3K/Akt/mTOR pathway: Effective combinations and clinical considerations. Drug Resist Updat. 11:32–50. 2008. View Article : Google Scholar : PubMed/NCBI
10	Martelli AM, Evangelisti C, Chiarini F, Grimaldi C, Manzoli L and McCubrey JA: Targeting the PI3K/AKT/mTOR signaling network in acute myelogenous leukemia. Expert Opin Investig Drugs. 18:1333–1349. 2009. View Article : Google Scholar : PubMed/NCBI
11	Ikezoe T, Nishioka C, Bandobashi K, Yang Y, Kuwayama Y, Adachi Y, Takeuchi T, Koeffler HP and Taguchi H: Longitudinal inhibition of PI3K/Akt/mTOR signaling by LY294002 and rapamycin induces growth arrest of adult T-cell leukemia cells. Leuk Res. 31:673–682. 2007. View Article : Google Scholar : PubMed/NCBI
12	Steelman LS, Abrams SL, Whelan J, Bertrand FE, Ludwig DE, Basecke J, Libra M, Stivala F, Milella M, et al: Contributions of the Raf/MEK/ERK, PI3K/PTEN/Akt/mTOR and Jak/STAT pathways to leukemia. Leukemia. 22:686–707. 2008. View Article : Google Scholar : PubMed/NCBI
13	Hotamisligil GS: Endoplasmic reticulum stress and the inflammatory basis of metabolic disease. Cell. 140:900–917. 2010. View Article : Google Scholar : PubMed/NCBI
14	Banhegyi G, Baumeister P, Benedetti A, Dong D, Fu Y, Lee AS, Li J, Mao C, Margittai E, Ni M, et al: Endoplasmic reticulum stress. Ann N Y Acad Sci. 1113:58–71. 2007. View Article : Google Scholar : PubMed/NCBI
15	Rahmani M, Davis EM, Crabtree TR, Habibi JR, Nguyen TK, Dent P and Grant S: The kinase inhibitor sorafenib induces cell death through a process involving induction of endoplasmic reticulum stress. Mol Cell Biol. 27:5499–5513. 2007. View Article : Google Scholar : PubMed/NCBI
16	Pae HO, Jeong SO, Jeong GS, Kim KM, Kim HS, Kim SA, Kim YC, Kang SD, Kim BN and Chung HT: Curcumin induces pro-apoptotic endoplasmic reticulum stress in human leukemia HL-60 cells. Biochem Biophys Res Commun. 353:1040–1045. 2007. View Article : Google Scholar : PubMed/NCBI
17	Haferlach T, Kohlmann A, Schnittger S, Dugas M, Hiddemann W, Kern W and Schoch C: Global approach to the diagnosis of leukemia using gene expression profiling. Blood. 106:1189–1198. 2005. View Article : Google Scholar : PubMed/NCBI
18	Song Q, Han CC, Xiong XP, He F, Gan W, Wei SH, Liu HH, Li L and Xu HY: PI3K-Akt-mTOR signal inhibition affects expression of genes related to endoplasmic reticulum stress. Genet Mol Res. 15:150378682016. View Article : Google Scholar
19	Kim HS, Kim TJ and Yoo YM: Melatonin combined with endoplasmic reticulum stress induces cell death via the PI3K/Akt/mTOR pathway in B16F10 melanoma cells. PLoS One. 9:e926272014. View Article : Google Scholar : PubMed/NCBI
20	Gutierrez A, Sanda T, Grebliunaite R, Carracedo A, Salmena L, Ahn Y, Dahlberg S, Neuberg D, Moreau LA, Winter SS, et al: High frequency of PTEN, PI3K, and AKT abnormalities in T-cell acute lymphoblastic leukemia. Blood. 114:647–650. 2009. View Article : Google Scholar : PubMed/NCBI
21	Kondo Y, Kanzawa T, Sawaya R and Kondo S: The role of autophagy in cancer development and response to therapy. Nat Rev Cancer. 5:726–734. 2005. View Article : Google Scholar : PubMed/NCBI
22	Di Nardo A, Kramvis I, Cho N, Sadowski A, Meikle L, Kwiatkowski DJ and Sahin M: Tuberous sclerosis complex activity is required to control neuronal stress responses in an mTOR-dependent manner. J Neurosci. 29:5926–5937. 2009. View Article : Google Scholar : PubMed/NCBI
23	Chen MH, Chiang KC, Cheng CT, Huang SC, Chen YY, Chen TW, Yeh TS, Jan YY, Wang HM, Weng JJ, et al: Antitumor activity of the combination of an HSP90 inhibitor and a PI3K/mTOR dual inhibitor against cholangiocarcinoma. Oncotarget. 5:2372–2389. 2014. View Article : Google Scholar : PubMed/NCBI
24	Pavlidou A and Vlahos NF: Molecular alterations of PI3K/Akt/mTOR pathway: A therapeutic target in endometrial cancer. ScientificWorldJournal. 2014:7097362014. View Article : Google Scholar : PubMed/NCBI
25	Bjornsti MA and Houghton PJ: The TOR pathway: A target for cancer therapy. Nat Rev Cancer. 4:335–348. 2004. View Article : Google Scholar : PubMed/NCBI
26	Qin L, Wang Z, Tao L and Wang Y: ER stress negatively regulates AKT/TSC/mTOR pathway to enhance autophagy. Autophagy. 6:239–247. 2010. View Article : Google Scholar : PubMed/NCBI
27	Chen XL, Fu JP, Shi J, Wan P, Cao H and Tang ZM: CXC195 induces apoptosis and endoplastic reticulum stress in human hepatocellular carcinoma cells by inhibiting the PI3K/Akt/mTOR signaling pathway. Mol Med Rep. 12:8229–8236. 2015. View Article : Google Scholar : PubMed/NCBI