Apelin‑13 induces autophagy in hepatoma HepG2 cells through ERK1/2 signaling pathway‑dependent upregulation of Beclin1

Huang,Qiulin; Liu,Xuan; Cao,Chao; Lei,Junyue; Han,Dong; Chen,Guodong; Yu,Jia; Chen,Linxi; Lv,Deguan; Li,Zhongyu

doi:10.3892/ol.2015.3991

Apelin‑13 induces autophagy in hepatoma HepG2 cells through ERK1/2 signaling pathway‑dependent upregulation of Beclin1

Authors:
- Qiulin Huang
- Xuan Liu
- Chao Cao
- Junyue Lei
- Dong Han
- Guodong Chen
- Jia Yu
- Linxi Chen
- Deguan Lv
- Zhongyu Li
View Affiliations

Affiliations: Department of General Surgery, First Affiliated Hospital of University of South China, Hengyang, Hunan 421001, P.R. China, Institute of Pharmacy and Pharmacology, University of South China, Hengyang, Hunan 421001, P.R. China, Department of Microbiology, School of Medicine, University of South China, Hengyang, Hunan 421001, P.R. China
Published online on: December 3, 2015 https://doi.org/10.3892/ol.2015.3991
Pages: 1051-1056

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 aim of the present study was to investigate the effect of Apelin‑13 on autophagy in hepatocellular carcinoma HepG2 cells and the underlying mechanism of the effect. The HepG2 cells were treated with Apelin‑13 at a final concentration of 0.0001, 0.001, 0.01 and 0.1 µmol/l for 24 h. Cells were also treated with 10 µmol/l PD98059 for 24 h. The expression of the extracellular signal‑regulated kinase (ERK)1/2, phosphorylated ERK1/2 (pERK1/2) and Beclin1 proteins were detected by western blot analysis. Beclin1 mRNA expression was also detected by reverse transcription‑polymerase chain reaction. Autophagy was observed using fluorescence microscopy subsequent to monodansylcadaverine (MDC) staining. Following treatment with the various concentrations of Apelin‑13, the expression of the ERK1/2 protein remained at a similar level, whereas the expression of pERK1/2 increased in a dose‑dependent manner. Compared with the control group, the increase was significant (P<0.05). Similarly, Beclin1 expression was upregulated at the protein and mRNA levels by Apelin‑13 treatment in a dose‑dependent manner and was significantly increased compared with the control group. However, following treatment with the Apelin‑13 inhibitor PD98059, the expression of pERK1/2, Beclin1 protein and Beclin1 mRNA were significantly decreased (P<0.05). In addition, Apelin‑13 induced the autophagy of HepG2 cells in a dose‑dependent manner, as revealed by MDC staining. PD98059 inhibited autophagy of HepG2 cells induced by Apelin‑13. Therefore, Apelin‑13 may promote autophagy in HepG2 cells by inducing the phosphorylation of ERK1/2 and upregulating the expression of Beclin1.

View Figures

View References

1	Meijer AJ and Codogno P: Regulation and role of autophagy in mammalian cells. Int J Biochem Cell Biol. 36:2445–2462. 2004. View Article : Google Scholar : PubMed/NCBI
2	Liang XH, Jackson S, Seaman M, Brown K, Kempkes B, Hibshoosh H and Levine B: Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature. 402:672–676. 1999. View Article : Google Scholar : PubMed/NCBI
3	Shoji-Kawata S, Sumpter R, Leveno M, Campbell GR, Zou Z, Kinch L, Wilkins AD, Sun Q, Pallauf K, MacDuff D, et al: Identification of a candidate therapeutic autophagy-inducing peptide. Nature. 494:201–206. 2013. View Article : Google Scholar : PubMed/NCBI
4	Furuya N, Yu J, Byfield M, Pattingre S and Levine B: The evolutionarily conserved domain of Beclin 1 is required for Vps34 binding, autophagy and tumor suppressor function. Autophagy. 1:46–52. 2005. View Article : Google Scholar : PubMed/NCBI
5	Pattingre S, Bauvy C and Codogno P: Amino acids interfere with the ERK1/2-dependent control of macroautophagy by controlling the activation of Raf-1 in human colon cancer HT-29 cells. J Biol Chem. 278:16667–16674. 2003. View Article : Google Scholar : PubMed/NCBI
6	Huang YC, Hung WC, Chen WT, Yu HS and Chai CY: Sodium arsenite-induced DAPK promoterhypermethylation and autophagy via ERK1/2 phosphorylation in humanuroepithelial cells. Chem Biol Interact. 181:254–262. 2009. View Article : Google Scholar : PubMed/NCBI
7	Tatemoto K, Hosoya M, Habata Y, Fujii R, Kakegawa T, Zou MX, Kawamata Y, Fukusumi S, Hinuma S, Kitada C, et al: Isolation and characterization of a novel endogenous peptide ligand for the human APJ receptor. Biochem Biophys Res Commun. 251:471–476. 1998. View Article : Google Scholar : PubMed/NCBI
8	Li F, Li L, Qin X, Pan W, Feng F, Chen F, Zhu B, Liao D, Tanowitz H, Albanese C, et al: Apelin-induced vascular smooth muscle cell proliferation: The regulation of cyclin D1. Front Biosci. 13:3786–3792. 2008. View Article : Google Scholar : PubMed/NCBI
9	Biederbick A, Kern HF and Elsässer HP: Monodansylcadaverine (MDC) is a specific in vivo marker for autophagic vacuoles. Eur J Cell Biol. 66:3–14. 1995.PubMed/NCBI
10	D'Alessandro LA, Meyer R and Klingmüller U: Hepatocellular carcinoma A systems biology perspective. Front Physiol. 4:282013. View Article : Google Scholar : PubMed/NCBI
11	He J, Gu D, Wu X, Reynolds K, Duan X, Yao C, Wang J, Chen CS, Chen J, Wildman RP, et al: Major causes of death among men and women in China. N Engl J Med. 353:1124–1134. 2005. View Article : Google Scholar : PubMed/NCBI
12	Klionsky DJ: The molecular machinery of autophagy: U nanswered questions. J Cell Sci. 118:7–18. 2005. View Article : Google Scholar : PubMed/NCBI
13	Massey AC, Zhang C and Cuervo AM: Chaperone-mediated autophagy in aging and disease. Curr Top Dev Biol. 73:205–235. 2006. View Article : Google Scholar : PubMed/NCBI
14	Wang ZH, Xu L, Duan ZL, Zeng LQ, Yan NH and Peng ZL: Beclin 1 mediated macroautophagy involves regulation of caspase-9 expression in cervical cancer HeLa cells. Gynecol Oncol. 107:107–113. 2007. View Article : Google Scholar : PubMed/NCBI
15	Duan ZL, Peng ZL and Wang ZH: Expression and involved sign alransduction pathway of autophagy gene Beclin 1 in epithelial ovarian cancer. Sichuan Da Xue Xue Bao Yi Xue Ban. 38:239–242. 2007.(In Chinese). PubMed/NCBI
16	Miracco C, Cosci E, Oliveri G, Luzi P, Pacenti L, Monciatti I, Mannucci S, De Nisi MC, Toscano M, Malagnino V, et al: Protein and mRNA expression of autophagy gene Beclin 1 in human brain tumours. Int J Oncol. 30:429–436. 2007.PubMed/NCBI
17	Roesly HB, Khan MR, Chen HD, Hill KA, Narendran N, Watts GS, Chen X and Dvorak K: The decreased expression of Beclin-1 correlates with progression to esophageal adenocarcinoma, The role of deoxycholic acid. Am J Physiol Gastrointest Liver Physiol. 302:G864–G872. 2012. View Article : Google Scholar : PubMed/NCBI
18	Zhou WH, Tang F, Xu J, Wu X, Yang SB, Feng ZY, Ding YG, Wan XB, Guan Z, Li HG, et al: Low expression of Beclin 1, associated with high Bcl-XL, predicts a malignant phenotype and poor prognosis of gastric cancer. Autophagy. 8:389–400. 2012. View Article : Google Scholar : PubMed/NCBI
19	Yang Z and Klionsky DJ: Eaten alive: A history of macroautophagy. Nat Cell Biol. 12:814–822. 2010. View Article : Google Scholar : PubMed/NCBI
20	Apel A, Zentgraf H, Büchler MW and Herr I: Autophagy - a double-edged sword in oncology. Int J Cancer. 125:991–995. 2009. View Article : Google Scholar : PubMed/NCBI
21	Lee SJ, Kim HP, Jin Y, Choi AM and Ryter SW: Beclin 1 deficiency is associated with increased hypoxia-induced angiogenesis. Autophagy. 7:829–839. 2011. View Article : Google Scholar : PubMed/NCBI