EGR‑1 plays a protective role in AMPK inhibitor compound C‑induced apoptosis through ROS‑induced ERK activation in skin cancer cells

Chuang,Kai-Cheng; Chen,Fan-Wen; Tsai,Meng-Hsiun; Shieh,Jeng-Jer

doi:10.3892/ol.2021.12565

April-2021 Volume 21 Issue 4

Full Size Image

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

An International Open Access Journal Devoted to General Medicine.

April-2021 Volume 21 Issue 4

Full Size Image

Article

EGR‑1 plays a protective role in AMPK inhibitor compound C‑induced apoptosis through ROS‑induced ERK activation in skin cancer cells

Authors:
- Kai-Cheng Chuang
- Fan-Wen Chen
- Meng-Hsiun Tsai
- Jeng-Jer Shieh
View Affiliations / Copyright

Affiliations: Institute of Biomedical Sciences, National Chung Hsing University, Taichung 402, Taiwan, R.O.C., Department of Management Information System, National Chung Hsing University, Taichung 402, Taiwan, R.O.C.
Article Number: 304
|
Published online on: February 19, 2021

https://doi.org/10.3892/ol.2021.12565
Expand metrics +

Abstract

Skin cancer is caused by abnormal proliferation, gene regulation and mutation of epidermis cells. Compound C is commonly used as an inhibitor of AMP‑activated protein kinase (AMPK), which serves as an energy sensor in cells. Recently, compound C has been reported to induce apoptotic and autophagic death in various skin cancer cell lines via an AMPK‑independent pathway. However, the signaling pathways activated in compound C‑treated cancer cells remain unclear. The present oligodeoxynucleotide‑based microarray screening assay showed that the mRNA expression of the zinc‑finger transcription factor early growth response‑1 (EGR‑1), which helps regulate cell cycle progression and cell survival, was significantly upregulated in compound C‑treated skin cancer cells. Compound C was demonstrated to induce EGR‑1 mRNA and protein expression in a time and dose‑dependent manner. Confocal imaging showed that compound C‑induced EGR‑1 protein expression was localized in the nucleus. Compound C was demonstrated to activate extracellular signal‑regulated kinase (ERK) phosphorylation. Inhibition of this compound C‑induced ERK phosphorylation downregulated the mRNA and protein expression of EGR‑1. In addition, removal of compound C‑induced reactive oxygen species (ROS) not only decreased ERK phosphorylation, but also inhibited compound C‑induced EGR‑1 expression. A functional assay showed that knock down of EGR‑1 expression in cancer cells decreased the survival rate while also increasing caspase‑3 activity and apoptotic marker expression after compound C treatment. However, no difference in autophagy marker light chain 3‑II protein expression was observed between compound C‑treated control cells and EGR‑1‑knockdown cells. Thus, it was concluded that that EGR‑1 may antagonize compound C‑induced apoptosis but not compound C‑induced autophagy through the ROS‑mediated ERK activation pathway.

View Figures

View References

1	Zhou G, Myers R, Li Y, Chen Y, Shen X, Fenyk-Melody J, Wu M, Ventre J, Doebber T, Fujii N, et al: Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Invest. 108:1167–1174. 2001. View Article : Google Scholar : PubMed/NCBI
2	Bae EJ, Cho MJ and Kim SG: Metformin prevents an adaptive increase in GSH and induces apoptosis under the conditions of GSH deficiency in H4IIE cells. J Toxicol Environ Health A. 70:1371–1380. 2007. View Article : Google Scholar : PubMed/NCBI
3	Vucicevic L, Misirkic M, Janjetovic K, Harhaji-Trajkovic L, Prica M, Stevanovic D, Isenovic E, Sudar E, Sumarac- Dumanovic M, Micic D and Trajkovic V: AMP-activated protein kinase-dependent and -independent mechanisms underlying in vitro antiglioma action of compound C. Biochem Pharmacol. 77:1684–1693. 2009. View Article : Google Scholar : PubMed/NCBI
4	Jin J, Mullen TD, Hou Q, Bielawski J, Bielawska A, Zhang X, Obeid LM, Hannun YA and Hsu YT: AMPK inhibitor compound C stimulates ceramide production and promotes Bax redistribution and apoptosis in MCF7 breast carcinoma cells. J Lipid Res. 50:2389–2397. 2009. View Article : Google Scholar : PubMed/NCBI
5	Meley D, Bauvy C, Houben-Weerts JH, Dubbelhuis PF, Helmond MT, Codogno P and Meijer AJ: AMP-activated protein kinase and the regulation of autophagic proteolysis. J Biol Chem. 281:34870–34879. 2006. View Article : Google Scholar : PubMed/NCBI
6	Vucicevic L, Misirkic M, Janjetovic K, Vilimanovich U, Sudar E, Isenovic E, Prica M, Harhaji-Trajkovic L, Kravic-Stevovic T, Bumbasirevic V and Trajkovic V: Compound C induces protective autophagy in cancer cells through AMPK inhibition-independent blockade of Akt/mTOR pathway. Autophagy. 7:40–50. 2011. View Article : Google Scholar : PubMed/NCBI
7	Jang JH, Lee TJ, Yang ES, Min do S, Kim YH, Kim SH, Choi YH, Park JW, Choi KS and Kwon TK: Compound C sensitizes Caki renal cancer cells to TRAIL-induced apoptosis through reactive oxygen species-mediated down-regulation of c-FLIPL and Mcl-1. Exp Cell Res. 316:2194–2203. 2010. View Article : Google Scholar : PubMed/NCBI
8	Milbrandt J: A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Science. 238:797–799. 1987. View Article : Google Scholar : PubMed/NCBI
9	Gashler A and Sukhatme VP: Early growth response protein 1 (Egr-1): Prototype of a zinc-finger family of transcription factors. Prog Nucleic Acid Res Mol Biol. 50:191–224. 1995. View Article : Google Scholar : PubMed/NCBI
10	Aicher WK, Sakamoto KM, Hack A and Eibel H: Analysis of functional elements in the human Egr-1 gene promoter. Rheumatol Int. 18:207–214. 1999. View Article : Google Scholar : PubMed/NCBI
11	Pagel JI and Deindl E: Early growth response 1-a transcription factor in the crossfire of signal transduction cascades. Indian J Biochem Biophys. 48:226–235. 2011.PubMed/NCBI
12	Yu J, Baron V, Mercola D, Mustelin T and Adamson ED: A network of p73, p53 and Egr1 is required for efficient apoptosis in tumor cells. Cell Death Differ. 14:436–446. 2007. View Article : Google Scholar : PubMed/NCBI
13	Yu J, de Belle I, Liang H and Adamson ED: Coactivating factors p300 and CBP are transcriptionally crossregulated by Egr1 in prostate cells, leading to divergent responses. Mol Cell. 15:83–94. 2004. View Article : Google Scholar : PubMed/NCBI
14	Yu J, Zhang SS, Saito K, Williams S, Arimura Y, Ma Y, Ke Y, Baron V, Mercola D, Feng GS, et al: PTEN regulation by Akt-EGR1-ARF-PTEN axis. EMBO J. 28:21–33. 2009. View Article : Google Scholar : PubMed/NCBI
15	Bae MH, Jeong CH, Kim SH, Bae MK, Jeong JW, Ahn MY, Bae SK, Kim ND, Kim CW, Kim KR and Kim KW: Regulation of Egr-1 by association with the proteasome component C8. Biochim Biophys Acta. 1592:163–167. 2002. View Article : Google Scholar : PubMed/NCBI
16	Kim HS, Hwang JT, Yun H, Chi SG, Lee SJ, Kang I, Yoon KS, Choe WJ, Kim SS and Ha J: Inhibition of AMP-activated protein kinase sensitizes cancer cells to cisplatin-induced apoptosis via hyper-induction of p53. J Biol Chem. 283:3731–3742. 2008. View Article : Google Scholar : PubMed/NCBI
17	Krones-Herzig A, Mittal S, Yule K, Liang H, English C, Urcis R, Soni T, Adamson ED and Mercola D: Early growth response 1 acts as a tumor suppressor in vivo and in vitro via regulation of p53. Cancer Res. 65:5133–5143. 2005. View Article : Google Scholar : PubMed/NCBI
18	Das A, Chendil D, Dey S, Mohiuddin M, Mohiuddin M, Milbrandt J, Rangnekar VM and Ahmed MM: Ionizing radiation down-regulates p53 protein in primary Egr-1^−/− mouse embryonic fibroblast cells causing enhanced resistance to apoptosis. J Biol Chem. 276:3279–3286. 2001. View Article : Google Scholar : PubMed/NCBI
19	Baron V, Adamson ED, Calogero A, Ragona G and Mercola D: The transcription factor Egr1 is a direct regulator of multiple tumor suppressors including TGFbeta1, PTEN, p53, and fibronectin. Cancer Gene Ther. 13:115–124. 2006. View Article : Google Scholar : PubMed/NCBI
20	Gitenay D and Baron VT: Is EGR1 a potential target for prostate cancer therapy? Future Oncol. 5:993–1003. 2009. View Article : Google Scholar : PubMed/NCBI
21	Yang SZ and Abdulkadir SA: Early growth response gene 1 modulates androgen receptor signaling in prostate carcinoma cells. J Biol Chem. 278:39906–39911. 2003. View Article : Google Scholar : PubMed/NCBI
22	Lu C, Shi Y, Wang Z, Song Z, Zhu M, Cai Q and Chen T: Serum starvation induces H2AX phosphorylation to regulate apoptosis via p38 MAPK pathway. FEBS Lett. 582:2703–2708. 2008. View Article : Google Scholar : PubMed/NCBI
23	Huang SW, Wu CY, Wang YT, Kao JK, Lin CC, Chang CC, Mu SW, Chen YY, Chiu HW, Chang CH, et al: p53 modulates the AMPK inhibitor compound C induced apoptosis in human skin cancer cells. Toxicol Appl Pharmacol. 267:113–124. 2013. View Article : Google Scholar : PubMed/NCBI
24	Datta R, Rubin E, Sukhatme V, Qureshi S, Hallahan D, Weichselbaum RR and Kufe DW: Ionizing radiation activates transcription of the EGR1 gene via CArG elements. Proc Natl Acad Sci USA. 89:10149–10153. 1992. View Article : Google Scholar : PubMed/NCBI
25	Kao JK, Wang SC, Ho LW, Huang SW, Lee CH, Lee MS, Yang RC and Shieh JJ: M2-like polarization of THP-1 monocyte-derived macrophages under chronic iron overload. Ann Hematol. 99:431–441. 2020. View Article : Google Scholar : PubMed/NCBI
26	Tsai MH, Wang HC, Lee GW, Lin YC and Chiu SH: A decision tree based classifier to analyze human ovarian cancer cDNA microarray datasets. J Med Syst. 40:212016. View Article : Google Scholar : PubMed/NCBI
27	Yang WL, Perillo W, Liou D, Marambaud P and Wang P: AMPK inhibitor compound C suppresses cell proliferation by induction of apoptosis and autophagy in human colorectal cancer cells. J Surg Oncol. 106:680–688. 2012. View Article : Google Scholar : PubMed/NCBI
28	Berasi SP, Huard C, Li D, Shih HH, Sun Y, Zhong W, Paulsen JE, Brown EL, Gimeno RE and Martinez RV: Inhibition of gluconeogenesis through transcriptional activation of EGR1 and DUSP4 by AMP-activated kinase. J Biol Chem. 281:27167–27177. 2006. View Article : Google Scholar : PubMed/NCBI
29	Liu DX, Qian D, Wang B, Yang JM and Lu Z: p300-Dependent ATF5 acetylation is essential for Egr-1 gene activation and cell proliferation and survival. Mol Cell Biol. 31:3906–3916. 2011. View Article : Google Scholar : PubMed/NCBI
30	Cao X, Mahendran R, Guy GR and Tan YH: Protein phosphatase inhibitors induce the sustained expression of the Egr-1 gene and the hyperphosphorylation of its gene product. J Biol Chem. 267:12991–12997. 1992. View Article : Google Scholar : PubMed/NCBI
31	Huang RP, Fan Y, deBelle I, Ni Z, Matheny W and Adamson ED: Egr-1 inhibits apoptosis during the UV response: Correlation of cell survival with Egr-1 phosphorylation. Cell Death Differ. 5:96–106. 1998. View Article : Google Scholar : PubMed/NCBI
32	Manente AG, Pinton G, Tavian D, Lopez-Rodas G, Brunelli E and Moro L: Coordinated sumoylation and ubiquitination modulate EGF induced EGR1 expression and stability. PLoS One. 6:e256762011. View Article : Google Scholar : PubMed/NCBI
33	Varshavsky A: The ubiquitin system. Trends Biochem Sci. 22:383–387. 1997. View Article : Google Scholar : PubMed/NCBI
34	Kyriakis JM, Banerjee P, Nikolakaki E, Dai T, Rubie EA, Ahmad MF, Avruch J and Woodgett JR: The stress-activated protein kinase subfamily of c-Jun kinases. Nature. 369:156–160. 1994. View Article : Google Scholar : PubMed/NCBI
35	Lim CP, Jain N and Cao X: Stress-induced immediate-early gene, egr-1, involves activation of p38/JNK1. Oncogene. 16:2915–2926. 1998. View Article : Google Scholar : PubMed/NCBI
36	Sarker KP and Lee KY: L6 myoblast differentiation is modulated by Cdk5 via the PI3K-AKT-p70S6K signaling pathway. Oncogene. 23:6064–6070. 2004. View Article : Google Scholar : PubMed/NCBI
37	Ha YM, Park EJ, Kang YJ, Park SW, Kim HJ and Chang KC: Valsartan independent of AT₁ receptor inhibits tissue factor, TLR-2 and −4 expression by regulation of Egr-1 through activation of AMPK in diabetic conditions. J Cell Mol Med. 18:2031–2043. 2014. View Article : Google Scholar : PubMed/NCBI
38	Birchmeier W and Birchmeier C: Epithelial-mesenchymal transitions in development and tumor progression. EXS. 74:1–15. 1995.PubMed/NCBI
39	Grotegut S, von Schweinitz D, Christofori G and Lehembre F: Hepatocyte growth factor induces cell scattering through MAPK/Egr-1-mediated upregulation of Snail. EMBO J. 25:3534–3545. 2006. View Article : Google Scholar : PubMed/NCBI
40	Vande Woude GF, Jeffers M, Cortner J, Alvord G, Tsarfaty I and Resau J: Met-HGF/SF: Tumorigenesis, invasion and metastasis. Ciba Found Symp. 212:119–132, 148-154. 1997.PubMed/NCBI
41	Harada T, Morooka T, Ogawa S and Nishida E: ERK induces p35, a neuron-specific activator of Cdk5, through induction of Egr1. Nat Cell Biol. 3:453–459. 2001. View Article : Google Scholar : PubMed/NCBI
42	Guillemot L, Levy A, Raymondjean M and Rothhut B: Angiotensin II-induced transcriptional activation of the cyclin D1 gene is mediated by Egr-1 in CHO-AT(1A) cells. J Biol Chem. 276:39394–39403. 2001. View Article : Google Scholar : PubMed/NCBI
43	Cabodi S, Morello V, Masi A, Cicchi R, Broggio C, Distefano P, Brunelli E, Silengo L, Pavone F, Arcangeli A, et al: Convergence of integrins and EGF receptor signaling via PI3K/Akt/FoxO pathway in early gene Egr-1 expression. J Cell Physiol. 218:294–303. 2009. View Article : Google Scholar : PubMed/NCBI
44	Mayr B and Montminy M: Transcriptional regulation by the phosphorylation-dependent factor CREB. Nat Rev Mol Cell Biol. 2:599–609. 2001. View Article : Google Scholar : PubMed/NCBI
45	Jhala US, Canettieri G, Screaton RA, Kulkarni RN, Krajewski S, Reed J, Walker J, Lin X, White M and Montminy M: cAMP promotes pancreatic beta-cell survival via CREB-mediated induction of IRS2. Genes Dev. 17:1575–1580. 2003. View Article : Google Scholar : PubMed/NCBI
46	Kang JH, Kim MJ, Ko SH, Jeong IK, Koh KH, Rhie DJ, Yoon SH, Hahn SJ, Kim MS and Jo YH: Upregulation of rat Ccnd1 gene by exendin-4 in pancreatic beta cell line INS-1: Interaction of early growth response-1 with cis-regulatory element. Diabetologia. 49:969–979. 2006. View Article : Google Scholar : PubMed/NCBI
47	Kang JH, Kim MJ, Jang HI, Koh KH, Yum KS, Rhie DJ, Yoon SH, Hahn SJ, Kim MS and Jo YH: Proximal cyclic AMP response element is essential for exendin-4 induction of rat EGR-1 gene. Am J Physiol Endocrinol Metab. 292:E215–E222. 2007. View Article : Google Scholar : PubMed/NCBI
48	León-Buitimea A, Rodriguez-Fragoso L, Lauer FT, Bowles H, Thompson TA and Burchiel SW: Ethanol-induced oxidative stress is associated with EGF receptor phosphorylation in MCF-10A cells overexpressing CYP2E1. Toxicol Lett. 209:161–165. 2012. View Article : Google Scholar : PubMed/NCBI
49	Lei H and Kazlauskas A: Growth factors outside of the platelet-derived growth factor (PDGF) family employ reactive oxygen species/Src family kinases to activate PDGF receptor alpha and thereby promote proliferation and survival of cells. J Biol Chem. 284:6329–6336. 2009. View Article : Google Scholar : PubMed/NCBI
50	Jeong SH, Park JH, Kim JN, Park YH, Shin SY, Lee YH, Kye YC and Son SW: Up-regulation of TNF-alpha secretion by cigarette smoke is mediated by Egr-1 in HaCaT human keratinocytes. Exp Dermatol. 19:e206–e212. 2010. View Article : Google Scholar : PubMed/NCBI
51	Kim JN, Kim HJ, Jeong SH, Kye YC and Son SW: Cigarette smoke-induced early growth response-1 regulates the expression of the cysteine-rich 61 in human skin dermal fibroblasts. Exp Dermatol. 20:992–997. 2011. View Article : Google Scholar : PubMed/NCBI
52	Thyss R, Virolle V, Imbert V, Peyron JF, Aberdam D and Virolle T: NF-kappaB/Egr-1/Gadd45 are sequentially activated upon UVB irradiation to mediate epidermal cell death. EMBO J. 24:128–137. 2005. View Article : Google Scholar : PubMed/NCBI
53	Cheong MW, Kuo LH, Cheng YN, Tsai PJ, Ho LC, Tai HC, Chiu WT, Chen SH, Lu PJ, Shan YS, et al: Loss of Egr-1 sensitizes pancreatic β-cells to palmitate-induced ER stress and apoptosis. J Mol Med (Berl). 93:807–818. 2015. View Article : Google Scholar : PubMed/NCBI
54	Parra E, Ferreira J and Ortega A: Overexpression of EGR-1 modulates the activity of NF-κB and AP-1 in prostate carcinoma PC-3 and LNCaP cell lines. Int J Oncol. 39:345–352. 2011.PubMed/NCBI