. 2020 May 5;35(1):1124–1136. doi: 10.1080/14756366.2020.1758691

Table 3.

Representative examples of cellular effects of EX-527.

Cell lines^a	Added agent	Effect of EX-527 on cells	Effect of EX-527 at the protein level	Comments	References
NCI-H460 U-2 OS MCF-7 HMEC	Etoposide, adriamycin, hydroxyurea, or hydrogen peroxide	No effect at 1 µM	Increases p53 acetylation (K382) at 1 µM (but no effect on two specific p53 target genes)	No effect on p53 without the genotoxic agent − 1 µM is non-toxic to all cell lines	Solomon et al.¹⁶
HCT-116	5-FU or camptothecin	Decreases cell proliferation and increases apoptosis at 2 µM	–	Increases cell proliferation at 2 µM, without the chemotherapy agent (and under growth factor deprivation)	Kabra et al.⁴⁰
MCF-7	None	Decreases proliferation at 50–100 µM	No apparent increase in p53 acetylation, but global increase in lysine acetylation of proteins	Causes cell cycle arrest at G1 phase at 50 µM	Peck et al.²⁰
U937	None	No cytotoxicity up to 50 µM ∼10 % apoptosis induction at 50 µM	–	No effect on granulocytic differentiation at 50 µM	Rotili at al. ²²
Primary AML Primary B-CLL U937 697 Jurkat	Valproic acid (VA): HDAC class I/II/IV inhibitor	Synergistic effect with VA (100 µg/mL): ∼60% leukaemia cell death at 75 µM	Effect through Bax: in Jurkat with increased Bax expression, ∼70% leukaemia cell death at 75 µM (even without VA)	Low cytotoxic activity in leukaemia cells without VA	Cea et al.⁴¹
SGC transfected with ATF4 (induces MDR effects)	5-FU or cisplatin	Increases the cytotoxicity of 5-FU and cisplatin at 10 µM (synergistic effect)	Downregulates MDR1 expression	Slightly increases the viability at 10 µM without the cytotoxic agent	Zhu et al.⁴²
MCF-7 U937	None	Cell cycle arrest in the G1 phase (no apoptosis) at 50 µM	At 10 µM, increases p53 and α-tubulin acetylation	No effect on granulocytic differentiation at 50 µM	Mellini et al.²³
CSC: CRC (CRO and 1.1) GBM (30P and 30PT)	None	Weak inhibition of cell viability at 50 µM (up to 20%)	–	In combination with SIRT2 inhibitor AGK2, slight synergic effect proposed	Rotili et al.⁴³
HCT-116	None	–	At 10 µM, increases p53 acetylation	Ratio (Ac-p53 / total p53) = 0.27 vs control = 0.03	Suzuki et al.⁴⁴
BMDMs	LPS-induced production of cytokines	At 4 µM, no effect on cytokine production by macrophages	–	No effect at 120 µM or in combination with SIRT2-selective inhibitors	Lugrin et al.⁴⁵
HCC (HepG2)	Trichostatin (TSA): HDAC inhibitor	–	At 20 µM: increases p53 acetylation decreases NAMPT enzymatic activity and increases its extracellular levels	–	Schuster et al.⁴⁶
PC-12 expressing mHtt	None	Rescues ∼35% mHtt mediated toxicity at 1 µM (but only ∼25% at 10 µM)	Increases mHtt acetylation and clearance	Protective effect in primary cultures of rat striatal neurons infected with viral vectors expressing a mHtt fragment	Smith et al.⁴⁷
SH-SY5Y	None	At 3 µM, restores viability in neuronal cells carrying a G93A SOD1 mutant (ALS-linked mutation)	No increase in p53 acetylation	The authors propose that the observed effects do not come from SIRT1 inhibition	Valle et al.⁴⁸
HUVEC	H₂O₂	At 15 µM, protects against H₂O₂: Increases cell viability, adhesion, migratory ability Decreases the apoptotic index and ROS production	Reverses H₂O₂ effects: Decreases SIRT1, p-JNK, p-p38MAPK and increases p-ERK expression	No effect on HUVEC untreated by H₂O₂	Li et al.⁴⁹
PANC-1 BXPC-3 ASPC-1	Gemcitabine or cisplatin	At 1 µM, increases the cytotoxic and pro-apoptotic effects of gemcitabine and cisplatin	At 2 µM, increases p53 acetylation and FOXO3a expression	Pro-apoptotic and anti-proliferative effects also without the cytotoxic agent (IC₅₀ values 5 to 9 µM)	Zhang et al.⁵⁰
TNBC MDA-MB-231 BT-549	None	Decreases viability by 20% at 50 µM	At 25 µM, increases p53 acetylation (K382)	Additional complex interplay with AMPK and metadherin studied	Gollavilli et al.⁵¹
CSCs: CD44^high CML K562 CD44⁺ HCT-15	Hsp90 inhibitors: 17-AAG and AUY922	At 10 nM, increases the cytotoxicity of Hsp90 inhibitors	Involvement of HSF1 and MDR related molecules proposed	–	Kim et al.⁵²
CEM/VLB₁₀₀ MCF7-MDR (MDR variants)	Hsp90 inhibitors: 17-AAG and AUY922	At 10 nM, increases the cytotoxicity of Hsp90 inhibitors (synergistic effect demonstrated)	At 50 nM: Decreases 17-AAG induced expression of Hsp70/Hsp27 Increases 17-AAG induced downregulation of mut p53 and P-gp Decreases P-gp efflux activity with AUY922	Decreases P-gp efflux activity also without AUY922	Kim et al.⁵³
HCC (HepG2)	H₂O₂	–	At 10 µM, aggravates H₂O₂ induced: Decrease in MnSOD and Bcl-xL Increase in cleaved caspase 3	–	Hu et al.⁵⁴
HHUA, HHUA-SIRT1, HEC151 and HEC1B	Cisplatin	At 1 µM, inhibits the proliferation with a synergic effect with cisplatin	Independent of p53 mutation status	Inhibits the proliferation at 1 µM also without cisplatin	Asaka et al.⁵⁵
Human platelets	None	At 10 µM, induces apoptosis-like changes: enhances annexin V binding, ROS production and drop in mitochondrial transmembrane potential	Increases p53 acetylation and the level of conformationally active Bax	–	Kumari et al.⁵⁶
Naïve CD4 T cells	None	At 12.5 µM, decreases Th17 effector cells differentiation from CD4 T cells	SIRT1 deacetylates RORγt and increases its transcriptional activity	–	Lim et al.⁵⁷
HeLa	None	At 10 µM, decreases colony formation (> 50 %) and migration At 50 µM, causes cell cycle arrest in the G1 phase (no apoptosis)	Increases HSF1 acetylation, ubiquitination, and degradation Causes G1 phase arrest mediated by inhibition of Cdk4, Cdk6 and cyclin D1	–	Kim et al.⁵⁸
Pluripotent P19 cells (mouse embryonic carcinoma)	None	At 100 µM, accelerates the differentiation of P19 cells into functionally active neurons	Identification of neuron-specific proteins and glutamate receptor in differentiated neurons	–	Kim et al.⁵⁹
A549	MK-1775: WEE1 kinase inhibitor (induces DNA damage)	At 5 µM, enhances the anti-proliferative and pro-apoptotic effects of MK-1775.	Reduces homologous recombination (HR) repair activity by acetylation of machinery proteins NBS1 and Rad51	Several other lung cancer cells lines tested give similar results	Chen et al.⁶⁰
THP-1 macrophages	Ox-LDL induced inhibition of autophagy	At 2 µM, increases the inhibition of autophagy	Exacerbates acetylation of Atg5	Macrophage accumulation is linked to atherosclerosis	Yang et al.⁶¹
AML12 RAW264.7 macrophages	[Ru(CO)₃Cl₂]₂ (Carbon monoxide releasing molecule)	At 10 µM, decreases the protective effect of [Ru(CO)₃Cl₂]₂ after hypoxia/reoxygenation injury	Decreases the inhibition of acetylation, translocation to the cytoplasm, and release of HMGB1 by [Ru(CO)₃Cl₂]₂	A direct deacetylation of HMGB1 by SIRT1 was also demonstrated with isolated enzymes	Sun et al.⁶²
U373 Hs683	None	Inhibits cell growth with IC₅₀ = 157.4 ± 23.0 (U373) and 115.9 ± 23.3 µM (Hs683)	–	–	Schnekenburger et al.³⁰
HCC (HepG2 and Huh7)	None	Decreases cell survival with IC₅₀ = 195 ± 12 (HepG2) and 33 ± 6 µM (Huh7) and increases early apoptosis at 1 µM	Increases p53 and FoxO1 acetylation at 1 µM Decreases ABC transporters P-gp and MRP3 protein levels at 40 µM in HepG2	3D cultures: decreases spheroid growth and viability with IC₅₀ = 567 ± 41 (HepG2) and 67 ± 16 µM (Huh7)	Ceballos et al.⁶³
T cells	None	At 50 µM, increases the number and the suppressive function of Tregs	Increases both the acetylation and the expression levels of FOXP3	T cells isolated from patients suffering from abdominal aortic aneurysm	Jiang et al.⁶⁴
HCC (HepG2)	Hesperetin	At 10 µM, no effect on cell viability	Inhibits the increase of SIRT1 activity and AMPK phosphorylation caused by hesperetin	–	Shokri Afra et al.⁶⁵
BMMs	RANKL-induced Osteoclastogenesis	Promotes RANKL-stimulated osteoclastogenesis	Increases TNF-α mRNA and protein levels and ROS production	Dose of EX-527 not found	Yan et al.⁶⁶
HUVEC	High glucose conditions Resveratrol	At 10 µM, abolishes resveratrol-mediated anti-apoptosis and pro-proliferation effects	Involvement of the transcription factors Foxo1 and c-Myc	–	Huang et al.⁶⁷
HL-7702	Isoniazid (antituberculosis drug)	At 1 µM, aggravates the cell damages caused by isoniazid	In combination with isoniazid, increases further the expression of inflammatory regulators and cytokines, and the level of H3K9 acetylation in the promoter region of the IL-6 gene	No effects on cells and proteins when used alone	Zhang et al.⁶⁸
T cells stimulated with allogenic APC (co-cultures)	None	At 10 µg/mL, reduces T cell proliferation	Increases p53 acetylation and total protein acetylation	–	Daenthanasanmak et al.⁶⁹
MDA-MB-231 (high NNMT expression)	Adriamycin or paclitaxel	Increases the cytotoxicity, the inhibition of colony formation, and the apoptosis caused by the cytotoxic agents	Decreases the protection against cytotoxic agents given by the high NNMT expression	No effect without a cytotoxic agent Dose of EX-527 not found	Wang et al.⁷⁰

^{^a}

Cell lines: 697: B cell precursor leukaemia; A549: adenocarcinomic human alveolar basal epithelial cells (lung cancer); AML12: alpha mouse liver 12 (from hepatocytes); ASPC-1: pancreatic cancer; B-CLL: B cell chronic lymphocytic leukaemia; BM(D)Ms: bone-marrow derived macrophages; BXPC-3: pancreatic cancer; CEM/VLB₁₀₀: MDR variant of acute lymphoblastic leukaemia cells (overexpressing P-gp); CML: human chronic leukaemia; CRC: colorectal cancer; CSCs: cancer stem-like cells; GBM: glioblastoma multiforme; HCC: hepatocellular carcinoma; HCT-116/HCT-15: human colon cancer; Hela: cervical cancer; HHUA, HEC151, and HEC1B: human endometrial carcinoma; HMEC: primary human mammary epithelial cells; HL-7702: human normal liver cells; Hs683: glioblastoma; HUVEC: human umbilical vein endothelial cells; Jurkat: acute T cell leukaemia; MCF-7: human breast cancer; MDA-MB-231: breast cancer; NCI-H460: human non-small cell lung cancer; PANC-1: pancreatic cancer; PC-12: rat pheochromocytoma cells; SGC7901: human gastric adenocarcinoma; SH-SY5Y: subclone from bone marrow cells from neuroblastoma; Th17: T helper 17 cells (not naïve CD4 T cells); THP-1: human leukaemia monocyte; TNBC: triple negative breast cancer; Tregs: T regulatory cells; U373: glioblastoma; U937: human myeloid leukaemia (AML: acute myelogenous leukaemia); U-2 OS: human bone osteosarcoma epithelial cells.

5-FU: 5-fluorouracil; ABC: ATP binding cassette; AMPK: AMP-activated protein kinase; APC: antigen-presenting cells; ATF4: activating transcription factor 4; Atg5: autophagy-related 5; Bcl-xL: B cell lymphoma-extra-large; FoxO: forkhead box O; FOXP3: human forkhead box P3; HMGB1: high-mobility group box 1; HSF1: heat shock factor 1; Hsp: heat shock protein; LPS: lipopolysaccharides; MRP3: multidrug resistance-associated protein 3; mHtt (mHttex1pQ72): mutated Htt (huntingtin) exon 1 fragment with expanded Q repeat, presenting aggregates, and cytotoxicity, model of Huntington’s disease (HD); MnSOD: manganese superoxide dismutase; NNMT: nicotinamide N-methyl transferase; Ox-LDL: oxidised low-density lipoprotein; P-gp/MDR1: P-glycoprotein/multidrug resistance protein 1; RANKL: receptor activator of nuclear factor-κB ligand; RORγt: RAR-related orphan receptor γ-t; TNF-α: tumour necrosis factor-α