Effects of various autophagy modulators on the expression of autophagic markers LC3II and p62

WEI Yanming; REN Jinhong; LUAN Zhihua; WANG Yonghui

doi:10.11665/j.issn.1000-5048.20180313

Journal of China Pharmaceutical University > 2018 > 49(3): 341-347. > DOI: 10.11665/j.issn.1000-5048.20180313

WEI Yanming, REN Jinhong, LUAN Zhihua, WANG Yonghui. Effects of various autophagy modulators on the expression of autophagic markers LC3II and p62[J]. Journal of China Pharmaceutical University, 2018, 49(3): 341-347. DOI: 10.11665/j.issn.1000-5048.20180313

Citation:

PDF (2598 KB)

Effects of various autophagy modulators on the expression of autophagic markers LC3II and p62

More Information

Graphical Abstract

Abstract

Abstract

Western blotting of autophagic markers LC3II and p62 are widely used for estimating autophagic activity. To compare the regulation of various autophagy modulators on LC3II and p62, HEK293 cells were treated separately with mTOR-dependent autophagy activator rapamycin or -independent autophagy activators trehalose, and autophagy inhibitors including 3-methyladenine(3-MA), bafilomycin A1 or E64d and pepstatin A that inhi-bited the initiation of autophagy, the fusion of autophagosome and lysosome, and the activities of lysosomal enzymes accordingly, and then LC3II and p62 levels were assessed. Western blot results demonstrated that rapamycin enhanced the conversion of LC3I to LC3II, promoted the degradation of p62 simultaneously, while trehalose merely increased the expression of LC3II with no influence on the p62 level. Moreover, inhibition of autophagy commonly led to accumulation of LC3II as well as blockage of p62 degradation in a concentration- and time- dependent manner. These results indicate that obvious differences exist in the regulation of LC3II and p62 by various modulators although both are autophagic markers.
- autophagy activator,
- autophagy inhibitor,
- LC3II,
- p62

FullText(HTML)

References (18)

References

[1]	Choi AM,Ryter SW,Levine B.Autophagy in human health and disease[J].N Engl J Med,2013,368(7):651-662.
[2]	Schneider JL,Cuervo AM.Autophagy and human disease:emerging themes[J].Curr Opin Genet Dev,2014,26:16-23.
[3]	Klionsky D,Abdalla F,Abeliovich H,et al.Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes[J].Autophagy,2012,8(4):445-544.
[4]	Klionsky D,Cuervo A,Seglen P.Methods for monitoring autophagy from yeast to human[J].Autophagy,2007,3(3):181-206.
[5]	Kabeya Y,Mizushima N,Ueno T,et al.LC3,a mammalian homologue of yeast Apg8p,is localized in autophagosome membranes after processing[J].EMBO J,2000,19(21):5720-5728.
[6]	Ichimura Y,Kominami E,Tanaka K,et al.Selective turnover of p62/A170/SQSTM1 by autophagy[J].Autophagy,2008,4(8):1063-1066.
[7]	Law BY,Mok SW,Wu AG,et al.New potential pharmacological functions of Chinese herbal medicines via regulation of autophagy[J].Molecules,2016,21(3):359.
[8]	Cheng Y,Ren X,Hait WN,et al.Therapeutic targeting of autophagy in disease:biology and pharmacology[J].Pharmacol Rev,2013,65(4):1162-1197.
[9]	Tanida I,Minematsu-Ikeguchi N,Ueno T,et al.Lysosomal turnover,but not a cellular level,of endogenous LC3 is a marker for autophagy[J].Autophagy,2005,1(2):84-91.
[10]	Kuusisto E,Suuronen T,Salminen A.Ubiquitin-binding protein p62 expression is induced during apoptosis and proteasomal inhibition in neuronal cells[J].Biochem Biophys Res Commun,2001,280(1):223-228.
[11]	Gao L, Lv G, Guo X, et al. Activation of autophagy protects against cholestasis-induced hepatic injury[J].Cell Biosci,2014,4:47.
[12]	Tang Q,Zheng G,Feng Z,et al.Trehalose ameliorates oxidative stress-mediated mitochondrial dysfunction and ER stress via selective autophagy stimulation and autophagic flux restoration in osteoarthritis development[J].Cell Death Dis,2017,8(10):e3081.
[13]	Wu Y,Wang X,Guo H,et al.Synthesis and screening of 3-MA derivatives for autophagy inhibitors[J].Autophagy,2013,9(4):595-603.
[14]	Yu M, Xu X, Jiang N, et al. Dehydropachymic acid decreases bafilomycin A1 induced β-amyloid accumulation in PC12 cells[J].J Ethnopharmacol,2017,198:167-173.
[15]	Gottlieb R,Andres A,Sin J,et al.Untangling autophagy measurements:all fluxed up[J].Circ Res,2015,116(3):504-514.
[16]	Mizushima N,Yoshimori T.How to interapret LC3 immunoblotting[J].Autophagy,2007,3(6):542-545.
[17]	Honma Y,Sato-Morita M,Katsuki Y,et al.Trehalose activates autophagy and decreases proteasome inhibitor-induced endoplasmic reticulum stress and oxidative stress-mediated cytotoxicity in hepatocytes[J].Hepatol Res,2018,48(1):94-105.
[18]	Lyu X,Hu Z.New methods to detect autophagic flux[J].Acta Pharm Sin(药学学报),2016,51(1):45-51.

Cited By

Get Citation

PDF

XML

Article views (2237) PDF downloads (4105)

Turn off MathJax

Article Contents

Abstract

References

Effects of various autophagy modulators on the expression of autophagic markers LC3II and p62

Abstract

References

Catalog

Export File

Citation

Format

Content