Ferritinophagy-Mediated ROS Production Contributed to Proliferation Inhibition, Apoptosis, and Ferroptosis Induction in Action of Mechanism of 2-Pyridylhydrazone Dithiocarbamate Acetate.

Longlong Li,Yun Fu,Shaoshan Li,Deng Guan,Yongli Li,Changzheng Li,Yalei Zhang,Jiankang Feng,Hao Li,Maria R Ciriolo

doi:10.1155/2021/5594059

Abstract

Reactive oxygen species (ROS) production is involved in the mechanism of action of a number of drugs, but the biological effects of ROS remain to be clarified. Furthermore, ferroptosis involves iron-dependent ROS production that may be derived from ferritinophagy; however, the association between ferroptosis and ferritinophagy has not been fully established. The present study demonstrated that dithiocarbamate derivatives (iron chelators) exhibited antineoplastic properties involving ferritinophagy induction, but whether the underlying mechanisms involved ferroptosis was unknown. To gain insight into the underlying mechanism, a dithiocarbamate derivative, 2-pyridylhydrazone dithiocarbamate s-acetic acid (PdtaA), was prepared. An MTT assay demonstrated that PdtaA inhibited proliferation involving ROS production (IC50 = 23.0 ± 1.5 μM for HepG2 cells). A preliminary mechanistic study revealed that PdtaA induced both apoptosis and cell cycle arrest. Notably, PdtaA also induced ferroptosis via downregulation of GPx4 and xCT, which was first reported for a dithiocarbamate derivative. Moreover, these cellular events were associated with ROS production. To explore the origin of ROS, expression of the ferritinophagy-related genes, ferritin, and nuclear receptor coactivator (NCOA4) were measured. Immunofluorescence and western blotting analysis indicated that PdtaA-induced ferritinophagy may contribute to ROS production. To investigate the role of ferritinophagy, autophagy inhibitor 3-methyladenin or genetic knockdown of NCOA4 was employed to inhibit ferritinophagy, which significantly neutralized the action of PdtaA in both apoptosis and ferroptosis. Taken together, PdtaA-induced cell cycle arrest, apoptosis, and ferroptosis were associated with ferritinophagy.

Highlights

Liver cancer is the third leading cause of deaths from cancer worldwide [1]
The new compound was purified by flash chromatography, and the structural characteristic of pyridylhydrazone dithiocarbamate s-acetic acid (PdtaA) was determined by NMR and HRMS spectra (Figure 1(a))
DNA breakage will stop DNA replication and trigger DNA repair mechanisms that promote the phosphorylation of the CDK2/Cyclin E complex at the S phase detection site, which leads to prolongation of the S phase [37, 38]

Summary

Introduction

Liver cancer is the third leading cause of deaths from cancer worldwide [1]. Chemotherapy is still an important clinical treatment, but its side effects and drug resistance make its therapeutic effect limited [2]. The regorafenib has been approved for treatment of advanced hepatocellular carcinoma; the treatment for liver cancer is not ideal; clearly, an effective therapeutic strategy still is required [3]. The studies should be focused on new molecularly targeted agents and assessing the benefits of combined molecular therapy under the heterogeneity condition of tumor [4]. The tumor microenvironment serves an important role in tumor metastasis [5], in particular metalloproteinases and cytokines degrade the extracellular matrix and recruit and suppress incoming immunocytes (such as monocyte), respectively. The recruited monocytes were differentiated and tamed to tumor-associated macrophages, which

Methods

Results

Conclusion