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Evaluation of the Effects of Serum Starvation and Hypoxic Conditions on Metabolic Pathway Protein Expressions in Breast and Hepatocellular Cancers
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Research Article
VOLUME: 72 ISSUE: 1
P: 39-48
April 2019

Evaluation of the Effects of Serum Starvation and Hypoxic Conditions on Metabolic Pathway Protein Expressions in Breast and Hepatocellular Cancers

J Ankara Univ Fac Med 2019;72(1):39-48
DOI: 10.4274/atfm.galenos.2019.40316
Gürcan Günaydın
Mustafa Emre Gedik
1. Hacettepe Üniversitesi Kanser Enstitüsü, Temel Onkoloji Anabilim Dalı, Ankara, Türkiye
No information available.
No information available
Received Date: 28.01.2019
Accepted Date: 12.04.2019
Publish Date: 13.06.2019
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ABSTRACT

Objectives:

Adenosine monophosphate (AMP) activating-protein-kinase (AMPK) is a crucial serine/threonine protein kinase that is activated in cellular metabolic stress conditions to maintain cellular energy-homeostasis. Under conditions of stress such as starvation and hypoxia, when the amount of energy in the organism is low or energy consumption is increased, the amount of cellular AMP increases and catabolic reactions such as fatty acid oxidation and glycolysis are induced due to AMPK activation. The most common conditions that cancer cells face in tumor-microenvironment were reported to be nutrient deficiency and lack of oxygen. This study aims to investigate whether cancer cells bear survival advantages with mechanisms of adaptation such as epithelial-mesenchymal-transition and metastasis by the regulation of AMPK under various stress conditions.

Materials and Methods:

p-AMPK, total-AMPK protein expression levels of breast (SKBR-3, MDA-MB-453) and hepatocellular cancer (HepG2, Huh-7) cell lines under conditions of serum starvation were determined by Western-blot method. In addition, HIF-1α, E-cadherin, p-AMPK and total-AMPK protein expression levels under conditions of hypoxia were also determined by Western-blot method. The effects of stress conditions on regulation of proteins associated with metabolic pathway and invasion in cancer cells were investigated. As a result, the in vitro effects of stress factors which are observed in tumor-microenvironment in vivo were experimentally investigated by this simulation approach.

Results:

Our findings demonstrate that conditions of starvation and hypoxia may show prominent and statistically significant effects on both cancer cell metabolic regulation and tumor progression. AMPK was shown to be regulated as a result of cellular stress in cancer cells (Huh7, SKBR3) with high AMPK gene expression. Such a regulation seems to provide an adaptive advantage to those cells.

Conclusion:

Our results will contribute to determining therapeutic strategies targeting cancer cell stress mechanisms and AMPK. In addition, the results will pave the way for future in vivo studies; thus, will contribute to the literature.

Keywords: AMPK, Hypoxia, Cellular Stress, Cancer Metabolism, Tumor Immunology

References

1
Proud CG. Regulation and roles of elongation factor 2 kinase. Biochem Soc Trans. 2015;43:328-332.
2
Carling D. AMPK signalling in health and disease. Curr Opin Cell Biol. 2017;45:31-37.
3
Frezza C, Zheng L, Tennant DA, et al. Metabolic profiling of hypoxic cells revealed a catabolic signature required for cell survival. PLoS One. 2011;6:e24411.
4
Russo GL, Russo M, Ungaro P. AMP-activated protein kinase: a target for old drugs against diabetes and cancer. Biochem Pharmacol. 2013;86:339-350.
5
Hardie DG, Ross FA, Hawley SA. AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nat Rev Mol Cell Biol. 2012;13:251-262.
6
Sanli T, Steinberg GR, Singh G, et al. AMP-activated protein kinase (AMPK) beyond metabolism: a novel genomic stress sensor participating in the DNA damage response pathway. Cancer Biol Ther. 2014;15:156-169.
7
Ross FA, MacKintosh C, Hardie DG. AMP-activated protein kinase: a cellular energy sensor that comes in 12 flavours. FEBS J. 2016;283:2987-3001.
8
Manning BD. Adaptation to starvation: translating a matter of life or death. Cancer Cell. 2013;23:713-715.
9
Dasgupta B, Chhipa RR. Evolving Lessons on the Complex Role of AMPK in Normal Physiology and Cancer. Trends Pharmacol Sci. 2016;37:192-206.
10
Hetz C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol. 2012;13:89-102.
11
Dejeans N, Manié S, Hetz C, et al. Addicted to secrete - novel concepts and targets in cancer therapy. Trends Mol Med. 2014;20:242-250.
12
Terai K, Hiramoto Y, Masaki M, et al. AMP-activated protein kinase protects cardiomyocytes against hypoxic injury through attenuation of endoplasmic reticulum stress. Mol Cell Biol. 2005;25:9554-9575.
13
Van Proeyen K, De Bock K, Hespel P. Training in the fasted state facilitates re-activation of eEF2 activity during recovery from endurance exercise. Eur J Appl Physiol. 2011;111:1297-1305.
14
Knight JR, Bastide A, Roobol A, et al. Eukaryotic elongation factor 2 kinase regulates the cold stress response by slowing translation elongation. Biochem J. 2015;465:227-238.
15
Valko M, Rhodes CJ, Moncol J, et al. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact. 2006;160:1-40.
16
Reuter S, Gupta SC, Chaturvedi MM, et al. Oxidative stress, inflammation, and cancer: how are they linked? Free Radic Biol Med. 2010;49:1603-1616.
17
Yang J, Zhang X, Zhang Y, et al. HIF-2a promotes epithelial-mesenchymal transition through regulating Twist2 binding to the promoter of E-cadherin in pancreatic cancer. J Exp Clin Cancer Res. 2016;35:26.
18
Zhang W, Shi X, Peng Y, et al. HIF-1a Promotes Epithelial-Mesenchymal Transition and Metastasis through Direct Regulation of ZEB1 in Colorectal Cancer. PLoS One. 2015;10:e0129603.
19
Pirkmajer S, Chibalin AV. Serum starvation: caveat emptor. Am J Physiol Cell Physiol. 2011;301:C272-9.
20
Wu D, Yotnda P. Induction and testing of hypoxia in cell culture. J Vis Exp. 2011;(54).
21
Sever R, Brugge JS. Signal transduction in cancer. Cold Spring Harb Perspect Med. 2015;5.
22
Gerlinger M, Rowan AJ, Horswell S, et al. Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med. 2012 Mar 8;366:883-892.
23
Giancotti FG. Deregulation of cell signaling in cancer. FEBS Lett. 2014 Aug 19;588:2558-2570.
24
Barretina J, Caponigro G, Stransky N, et al. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature. 2012;483:603-607.
25
Laderoute KR, Calaoagan JM, Chao WR, et al. 5’-AMP-activated protein kinase (AMPK) supports the growth of aggressive experimental human breast cancer tumors. J Biol Chem. 2014;289:22850-22864.
26
Li W, Saud SM, Young MR, et al. Targeting AMPK for cancer prevention and treatment. Oncotarget. 2015;6:7365-7378.
27
Techasen A, Loilome W, Namwat N, et al. Loss of E-cadherin promotes migration and invasion of cholangiocarcinoma cells and serves as a potential marker of metastasis. Tumour Biol. 2014;35:8645-8652.
28
Gheldof A, Berx G. Cadherins and epithelial-to-mesenchymal transition. Prog Mol Biol Transl Sci. 2013;116:317-336.
29
Kalluri R, Weinberg RA. The basics of epithelial-mesenchymal transition. J Clin Invest. 2009;119:1420-1428.
30
Carrizo M. Loss of E-Cadherin Expression and Epithelial Mesenchymal Transition (EMT) as Key Steps in Tumor Progression. J Cancer Prev Curr Res. 2017;7.
31
Saxena M, Balaji SA, Deshpande N, et al. AMP-activated protein kinase promotes epithelial-mesenchymal transition in cancer cells through Twist1 upregulation. J Cell Sci. 2018;131.
32
Li N, Huang D, Lu N, et al. Role of the LKB1/AMPK pathway in tumor invasion and metastasis of cancer cells (Review). Oncol Rep. 2015;34:2821-2826.
33
Kim H, Lin Q, Glazer PM, et al. The hypoxic tumor microenvironment in vivo selects the cancer stem cell fate of breast cancer cells. Breast Cancer Res. 2018;20:16.
34
Wu M, Swartz MA. Modeling tumor microenvironments in vitro. J Biomech Eng. 2014;136:021011.
35
Salvatore V, Teti G, Focaroli S, et al. The tumor microenvironment promotes cancer progression and cell migration. Oncotarget. 2017;8:9608-9616.
36
Binnewies M, Roberts EW, Kersten K, et al. Understanding the tumor immune microenvironment (TIME) for effective therapy. Nat Med. 2018;24:541-550.
37
Noman MZ, Hasmim M, Messai Y, et al. Hypoxia: a key player in antitumor immune response. A Review in the Theme: Cellular Responses to Hypoxia. Am J Physiol Cell Physiol. 2015;309:C569-79.
38
Blagih J, Coulombe F, Vincent EE, et al. The energy sensor AMPK regulates T cell metabolic adaptation and effector responses in vivo. Immunity. 2015;42:41-54.
39
Bai A, Ma AG, Yong M, et al. AMPK agonist downregulates innate and adaptive immune responses in TNBS-induced murine acute and relapsing colitis. Biochem Pharmacol. 2010;80:1708-1717.
40
Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144:646-674.
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