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Inhibition of SIRT1/2 upregulates HSPA5 acetylation and induces pro-survival autophagy via ATF4-DDIT4-mTORC1 axis in human lung cancer cells

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Abstract

Sirtuins have emerged as a promising novel class of anti-cancer drug targets. Inhibition of SIRT1 and SIRT2 induces apoptosis in cancer cells and they play multifaceted roles in regulating autophagy. In the present study, we found that salermide, a SIRT1/2-specific inhibitor or small interfering RNAs (siRNAs) to block SIRT1/2 expression could induce autophagy in human NSCLC cells. Moreover, SIRT1/2 inhibition increased the expression levels of ATF4 and DDIT4 and downregulated p-RPS6KB1 and p-EIF4EBP1, two downstream molecules of mTORC1. Moreover, ATF4 or DDIT4 knockdown attenuated salermide-induced autophagy, suggesting that SIRT1/2 inhibition induced autophagy through the ATF4-DDIT4-mTORC1 axis. Mechanistically, SIRT1/2 inhibition led to HSPA5 acetylation and dissociation from EIF2AK3, leading to ER stress response and followed by upregulation of ATF4 and DDIT4, triggering autophagy. Silencing of the autophagic gene ATG5 in lung cancer cells resulted in increased apoptotic cell death induced by SIRT1/2 inhibition. Our data show that inhibition of SIRT1/2 induces pro-survival autophagy via acetylation of HSPA5 and subsequent activation of ATF4 and DDIT4 to inhibit the mTOR signaling pathway in NSCLC cells. These findings suggest that combinatorial treatment with SIRT1/2 inhibitors and pharmacological autophagy inhibitors is an effective therapeutic strategy for cancer therapy.

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Abbreviations

NSCLC:

Non-small cell lung cancer

SLM:

Salermide

MAP1LC3B:

Microtubule-associated proteins 1A/1B light chain 3B

mTOR:

Mechanistic target of rapamycin

mTORC1:

Mechanistic target of rapamycin complex 1

UPR:

Unfolded protein response

PI3K:

Phosphoinositide-3 kinase

RPS6KB1:

70-kDa ribosomal protein S6 kinase beta-1

EIF4EBP1:

Eukaryotic translation initiation factor 4E-binding protein 1

ATF4:

Activating transcription factor 4

DDIT4:

DNA-damage-inducible transcript 4

SIRT1:

Sirtuin 1

SIRT2:

Sirtuin 2

HSPA5:

Heat shock 70-kDa protein 5

EIF2AK3:

Eukaryotic translation initiation factor 2-alpha kinase 3

SBD:

Substrate binding domain

CASP3:

Caspase 3: apoptosis-related cysteine peptidase

CASP8:

Caspase 8: apoptosis-related cysteine peptidase

PARP1:

Poly (ADP-ribose) polymerase 1

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Acknowledgements

This work was supported by grants from National Natural Science Foundation of China (81672855, 31571422, 31771526, 81472686) and Shandong Provincial Key Laboratory of Animal Cell and Developmental Biology (SPKLACDB-2019017 and SPKLACDB-2019012). We thank Haiyan Yu, Xiaomin Zhao and Sen Wang from SKLMT (State Key Laboratory of Microbial Technology, Shandong University) for the assistance in microimaging of LSCM analysis.

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Authors and Affiliations

  1. Shandong Provincial Key Laboratory of Animal Cell and Developmental Biology, School of Life Sciences, Shandong University, Qingdao, People’s Republic of China

    Ning Mu, Yuanjiu Lei, Yu Wang, Yingying Wang, Qinghui Duan, Guilin Ma, Xiangguo Liu & Ling Su

  2. Shandong Provincial Collaborative Innovation Center of Cell Biology, School of Life Sciences, Shandong Normal University, Jinan, People’s Republic of China

    Ning Mu, Xiangguo Liu & Ling Su

  3. Taishan College, Shandong University, Qingdao, People’s Republic of China

    Qinghui Duan

  4. Shandong University School of Life Science, Room N8-108, 72 Binhai Road, Qingdao, 266237, People’s Republic of China

    Ling Su

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  1. Ning Mu
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Mu, N., Lei, Y., Wang, Y. et al. Inhibition of SIRT1/2 upregulates HSPA5 acetylation and induces pro-survival autophagy via ATF4-DDIT4-mTORC1 axis in human lung cancer cells. Apoptosis 24, 798–811 (2019). https://doi.org/10.1007/s10495-019-01559-3

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