Proteomic analysis revealed association of aberrant ROS signaling with suberoylanilide hydroxamic acid-induced autophagy in Jurkat T-leukemia cells
Authors: Jingyi Li
Quek Choon Lau
Shengtao Zhou
Na Xie
Chunlai Nie
Yunlong Lei
Canhua Huang
Lijuan Chen
Rui Liu
Kui Wang
Yuquan Wei
Journal: Autophagy (2010)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: SAHA; autophagy; Jurkat T-leukemia cells; LC3-II; Beclin 1
Affiliations: The State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, P.R. China.

Abstract:  Suberoylanilide hydroxamic acid (SAHA) is a newly emerging histone deacetylase inhibitor (HDACi) and has been approved in phase II clinical trials for treating patients with cutaneous T-cell lymphoma. Autophagy is a conserved self-digestion process that degrades cytoplasmic materials and recycles long-lived proteins and organelles within cells. In this study, we demonstrate that SAHA stimulates autophagy in Jurkat T-leukemia cells, which was evidenced by the appearance of autophagic vacuoles, formation of acidic vesicular organelles, recruitment of LC3-II to the autophagosomes and conversion of LC3-I to LC3-II . Moreover, SAHA treatment upregulated expression of Beclin 1 and Atg7 and promoted formation of the Atg12-Atg5 conjugate. Furthermore, inhibition of autophagy by chloroquine (CQ) enhanced SAHA-induced apoptosis. To determine the underlying mechanism of SAHA-induced autophagy, two complementary proteomic approaches (2-DE and SILAC), coupled with ESI-Q-TOF MS/MS analysis are utilized to profile differentially expressed proteins between control and SAHA-treated Jurkat T-leukemia cells. In total, 72 proteins were identified with significant alterations. Cluster analysis of the changed proteins reveal several groups of enzymes associated with energy metabolism, anti-oxidative stress and cellular redox control, which suggested an abnormal reactive oxygen species (ROS) production in SAHA-treated Jurkat T-leukemia cells. These observations were further confirmed by ROS chemiluminescence assay. Mechanistic studies revealed that SAHA-triggered autophagy was mediated by ROS production, which could be attenuated by N-acetyl cysteine (NAC), a ROS inhibitor. Finally, we illustrated that Akt-mTOR signaling, a major suppressive cascade of autophagy, was inactivated by SAHA treatment. Taken together, our study identifies autophagy as a reaction to counter increased ROS and is thus involved as a cellular prosurvival mechanism in response to SAHA treatment. 

Retracted: No

PubPeer comments:
 Figure 2A appears to show an image overlap with different treatments (0.1 u03bcM TsA and 2.5 u03bcM sAhA), and different dimensions. *** Inverted ![file] *** Original ![file] *** Full figure ![file] , 
 #1 Thanks for pointing out the erroru00a0in our publication. Afteru00a0carefully examiningu00a0the data, weu00a0found that the mistake in Figure 2A was inadvertently made during manuscript preparation. We have contacted the journal editors to correct the error.u00a0Thanks again for your comment. Jingyi Li , 
 #2 Thank you for your reply. Could the authors please post the corrected figure here? , 
 Corrigendum, Published online: 12 Feb 2020 In Figure 2A, the image of TSA-treated Jurkat T-leukemia cells (24 h) was inadvertently misplaced with an image of SAHA-treated Jurkat T-leukemia cells (48 h) during figure assembly. Figure 2A should be corrected as below, and this correction do not change the description, interpretation, or the original conclusions of the manuscript. The authors apologize for any inconvenience caused. ![file] ,