Integrative proteomic and functional analyses provide novel insights into the action of the repurposed drug candidate nitroxoline in AsPC-1 cells

Abstract

We recently identified nitroxoline as a repurposed drug candidate in pancreatic cancer (PC) showing a dose-dependent antiproliferative activity in different PC cell lines. This antibiotic is effective in several in vitro and animal cancer models. To date, the mechanisms of nitroxoline anticancer action are largely unknown. Using shotgun proteomics we identified 363 proteins affected by nitroxoline treatment in AsPC-1 pancreatic cancer cells, including 81 consistently deregulated at both 24-and 48-hour treatment. These proteins previously unknown to be affected by nitroxoline were mostly downregulated and interconnected in a single highly-enriched network of protein-protein interactions. Integrative proteomic and functional analyses revealed nitroxoline-induced downregulation of Na/K-Atpase pump and β-catenin, which associated with drastic impairment in cell growth, migration, invasion, increased ROS production and induction of DNA damage response. Remarkably, nitroxoline induced a previously unknown deregulation of molecules with a critical role in cell bioenergetics, which resulted in mitochondrial depolarization. Our study also suggests that deregulation of cytosolic iron homeostasis and of co-translational targeting to membrane contribute to nitroxoline anticancer action. This study broadens our understanding of the mechanisms of nitroxoline action, showing that the drug modulates multiple proteins crucial in cancer biology and previously unknown to be affected by nitroxoline.

Nitroxoline inhibits bladder cancer progression by reversing EMT process and enhancing anti-tumor immunity

“…It has been reported that nitroxoline results in the suppression of Na/K-ATPase pump and β-catenin, which is closely related with cell growth, migration, invasion, Reactive Oxygen Species (ROS) generation and DNA damage response [5]. Our study demonstrated that nitroxoline led to significant inhibition of cell viability and induced apoptosis of bladder cancer cells in vitro.…”

“…The current research shows that nitroxoline has a significant inhibitory effect on many solid tumors, such as myeloma, breast cancer, glioma, pancreatic cancer, and prostate cancer, etc. [4][5][6][7][8][9][10][11], but its effect on bladder cancer is poorly known. Multiple mechanisms have been described as possible modes of the antitumor action of nitroxoline, including the activation of cell apoptosis [6], downregulation of Na/K-ATPase pump and β-catenin [5], and inhibition…”

Synthesis and evaluation of a large library of nitroxoline derivatives as pancreatic cancer antiproliferative agents

“…In this respect, it has been reported that several non-anticancer agents approved for the treatment of different human diseases may have anticancer properties 4 . Our recent studies provided the first evidence that the non-antitumor drug nitroxoline has antitumor effects on PC cells 5,6 . Previously, this antibiotic showed antitumour effects in preclinical cancer models including xenograft and genetically modified mice models of glioblastoma, myeloma, fibrosarcoma as well as bladder, prostate, kidney and breast cancers [7][8][9][10][11][12][13] .…”

“…In line with the results obtained in other tumours, we showed that nitroxoline significantly reduces cell viability, affects cell cycle, promotes apoptosis and markedly decreases clonogenic activity in PC cells 5 . Moreover, using integrative proteomic and functional approaches, we evaluated nitroxoline effects on protein expression in PC cells, showing that this drug affects several biological pathways and oncogenic proteins, previously unknown to play a role in nitroxoline anticancer activity 6 . Notably, our study revealed a pleiotropic mechanism of nitroxoline anticancer action that involves processes playing a crucial role in growth, migration, invasion and cell death, ROS production and DNA damage 6 .…”

“…Moreover, using integrative proteomic and functional approaches, we evaluated nitroxoline effects on protein expression in PC cells, showing that this drug affects several biological pathways and oncogenic proteins, previously unknown to play a role in nitroxoline anticancer activity 6 . Notably, our study revealed a pleiotropic mechanism of nitroxoline anticancer action that involves processes playing a crucial role in growth, migration, invasion and cell death, ROS production and DNA damage 6 . Remarkably, this drug induced a previously unknown deregulation of molecules that are involved in cell bioenergetics, leading to mitochondrial depolarisation 6 .…”