Doxorubicin metabolism moderately attributes to putative toxicity in prodigiosin/doxorubicin synergism in vitro cells

Shian-Ren Lin,Ching-Cheng Chen,Tsung-Jui Wu,Lebin Weng,Feng-Jen Tseng,Ching-Feng Weng,Chun-Shu Lin

doi:10.1007/s11010-020-03864-x

Abstract

Doxorubicin (Dox) is a widely neoplasm chemotherapeutic drug with high incidences of cardiotoxicity. Prodigiosin (PG), a red bacterial pigment from Serratia marcescens, has been demonstrated to potentiate Dox’s cytotoxicity against oral squamous cell carcinoma cells through elevating Dox influx and identified as a Dox enhancer via PG-induced autophagy; however, toxicity of normal cell remains unclear. This study is conducted to evaluate putative cytotoxicity features of PG/Dox synergism in the liver, kidney, and heart cells and further elucidate whether PG augmented Dox’s effect via modulating Dox metabolism in normal cells. Murine hepatocytes FL83B, cardio-myoblast h9c2, and human kidney epithelial cells HK-2 were sequentially treated with PG and Dox by measuring cell viability, cell death characteristics, oxidative stress, Dox flux, and Dox metabolism. PG could slightly significant increase Dox cytotoxicity in all tested normal cells whose toxic alteration was less than that of oral squamous carcinoma cells. The augmentation of Dox cytotoxicity might be attributed to the increase of Dox-mediated ROS accumulation that might cause slight reduction of Dox influx and reduction of Dox metabolism. It was noteworthy to notice that sustained cytotoxicity appeared in normal cells after PG and Dox were removed. Taken together, moderately metabolic reduction of Dox might be ascribed to the mechanism of increase Dox cytotoxicity in PG-induced normal cells; nevertheless, the determination of PG/Dox dose with sustained cytotoxicity in normal cells needs to be comprehensively considered.

Highlights

Rendering non-tissue specific characteristics, doxorubicin (Dox) has wide indications, e.g., leukemia, neuroblastoma, breast, and ovarian carcinoma, and most recurrent or metastatic cancer in cancer regimen [1], and it has severe adverse effects in cancer patients. 31–51% of Dox is excreted via biliary tract in their original form and < 10% of Dox metabolite is eliminated via urine after intake [2, 3]
Three pathways of Dox metabolism are under investigation: one-electron reduction, two-electron reduction, and deglycosylation, which carried about 50% of Dox out of cells [4]
Three cell lines were cultured in different mediums and were all supplied with 10% fetal bovine serum and 1% antibiotic/antimycotic and 0.2% normocinTM: FL83B in Kaighn-modified Ham’s F12 medium (F12K), h9c2 in Dulbecco’s modified Eagle medium (DMEM), and HK-2 in DMEM/F12 with additional 0.5 ng/ mL epidermal growth factor (EGF)

Summary

Introduction

Rendering non-tissue specific characteristics, doxorubicin (Dox) has wide indications, e.g., leukemia, neuroblastoma, breast, and ovarian carcinoma, and most recurrent or metastatic cancer in cancer regimen [1], and it has severe adverse effects in cancer patients. 31–51% of Dox is excreted via biliary tract in their original form and < 10% of Dox metabolite is eliminated via urine after intake [2, 3]. 31–51% of Dox is excreted via biliary tract in their original form and < 10% of Dox metabolite is eliminated via urine after intake [2, 3]. Three pathways of Dox metabolism are under investigation: one-electron reduction, two-electron reduction, and deglycosylation, which carried about 50% of Dox out of cells (remaining 50% will be excreted from their original form) [4]. One-electron reduction occurs in mitochondria which operates by NADH-ubiquinone oxidoreductase (NDUFS), xanthine oxidase (XOD), and a cytosolic enzyme NADPH: quinone oxidoreductase (NQO). The end-product of oneelectron reduction is Dox-semiquinone, whose ROS activity is the main cause of oxidative stress. Along with the trigger mechanism, the cytotoxicity of Dox is considered to be via two approaches: apoptotic activation, which refers to hematopoietic inhibition, mucositis, and enteritis; and oxidative stress which causes most hepatotoxicity, nephrotoxicity, and cardiotoxicity [8, 9]

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