Metabolomics profiles delineate uridine deficiency contributes to mitochondria-mediated apoptosis induced by celastrol in human acute promyelocytic leukemia cells

Abstract

Celastrol, extracted from “Thunder of God Vine”, is a promising anti-cancer natural product. However, its effect on acute promyelocytic leukemia (APL) and underlying molecular mechanism are poorly understood. The purpose of this study was to explore its effect on APL and underlying mechanism based on metabolomics. Firstly, multiple assays indicated that celastrol could induce apoptosis of APL cells via p53-activated mitochondrial pathway. Secondly, unbiased metabolomics revealed that uridine was the most notable changed metabolite. Further study verified that uridine could reverse the apoptosis induced by celastrol. The decreased uridine was caused by suppressing the expression of gene encoding Dihydroorotate dehydrogenase, whose inhibitor could also induce apoptosis of APL cells. At last, mouse model confirmed that celastrol inhibited tumor growth through enhanced apoptosis. Celastrol could also decrease uridine and DHODH protein level in tumor tissues. Our in vivo study also indicated that celastrol had no systemic toxicity at pharmacological dose (2 mg/kg, i.p., 21 days). Altogether, our metabolomics study firstly reveals that uridine deficiency contributes to mitochondrial apoptosis induced by celastrol in APL cells. Celastrol shows great potential for the treatment of APL.

Metabolomics Reveals that Cysteine Metabolism Plays a Role in Celastrol-Induced Mitochondrial Apoptosis in HL-60 and NB-4 Cells

“…However, HL-60 can respond to ATRA 22 , which is widely used as a cell line in the APL studies [24][25][26][27] . In our previous study 28 , consistent to previous reports 15,[19][20][21] , we also found celastrol caused apoptosis in HL-60 cells, indicating the key role of apoptosis in the effect of celastol in the treatment of acute leukemia.…”

“…Surveying these small molecules provides an overall understanding of biological mechanisms, thereby creating a more complete picture of the phenotype (the observable characteristics of a living system). In our previous study, we used metabolomics to study the underlying mechanism in HL-60 cells in vitro, and identified uridine was the most notable changed metabolite after celastrol treatment 28 . However, the metabolome changes in vivo are unknown.…”

Effect and mechanisms of celastrol on the apoptosis of HOS osteosarcoma cells

“…To maintain homeostasis of HOS cells, a large number of unfolded proteins activate the PERK signaling pathway by binding to Bip, and PERK, which is an important transducer, activates downstream molecular partners and protein secretion related to molecular gene transcription. To explore whether attenuating the expression of PERK signaling pathway members would reverse the protective effect of ERS induced by celastrol and further promote the apoptosis of HOS cells, we used an inhibitor of PERK (GSK2656157, 5 mM) to pretreat HOS cells for 1 h before celastrol treatment (19,20). Following treatment with GSK2656157, the proliferation of HOS cells was further inhibited compared with that of the celastrol group; karyopyknosis, condensation, karyorrhexis and other typical apoptotic bodies were more evident; and the apoptosis rates further increased, while immunofluorescence revealed that the expression of p-PERK was significantly attenuated (Fig.…”

“…In addition, numerous studies have revealed that many components extracted from traditional Chinese medicine have antitumor effects (3,8,10). Celastrol, extracted from the root of T. wilfordii, has been revealed to inhibit the proliferation of tumors, and the mitochondrial apoptotic pathway and death receptor apoptosis pathway were involved in the celastrol-induced apoptosis (19,20). Research has reported the effect of celastrol on osteosarcoma, which has demonstrated great antitumor potential.…”

Application of Metabolomics in the Study of Natural Products

“…Metabolomics can be used to evaluate hepatotoxicity [ 47 ], cerebral lesion [ 48 ], and nephrotoxicity [ 49 ], and evaluate the safety of natural products, such as noscapine and Butea monosperma extract (Table 1 ) [ 50 , 51 ]. Using the powerful technology, the toxicity and safety of various natural products have been evaluated, such as noscapine [ 50 ], bupleurotoxin [ 48 ], celastrol [ 52 ], TP [ 5 ], and various extracts of natural products (Table 1 and Fig. 8 ).…”