Comparative Mitochondrial Proteomic Analysis of Raji Cells Exposed to Adriamycin

Abstract

The antitumor mechanisms of adriamycin (ADR) have been thought to contribute to induction of apoptosis and inefficiency of DNA repair, processes that are to a large extent mediated by mitochondria. This study aimed to investigate characteristics of ADR, including its antineoplastic activity, drug resistance, and unexpected toxicity in non-Hodgkin lymphoma (NHL) Raji cells at the mitochondrial proteomic level. The alterations of the mitochondrial proteome of Raji cells treated with ADR were analyzed by twodimensional differential in-gel electrophoresis (2D-DIGE) coupled with linear ion trap quadrupole-electrospray ionization tandem mass spectrometry (LTQ-ESI-MS/MS). The altered patterns of three identified proteins were validated by Western blot and analyzed by pathway studio software. The results showed that 34 proteins were downregulated and 3 proteins upregulated in the study group compared with the control group. The differentially expressed proteins distributed their functions in reduction-oxidation reactions, DNA repair, cell cycle regulation, transporters and channels, and oxidative phosphorylation. Furthermore, heat shock protein 70 (HSP70), ATP-binding cassette transporter isoform B6 (ABCB6), and prohibitin (PHB) identified in this study may be closely related to chemoresistance and could serve as potential chemotherapeutic targets for NHL. Collectively, these results suggest that specific mitochondrial proteins are uniquely susceptible to alterations in abundance following exposure to ADR and carry implications for the investigation of therapeutic and prognostic markers. Further studies focusing on these identified proteins will be used to predict treatment response and reverse apoptosis resistance, and to explore drug-combination strategies associated with ADR for NHL therapy.

Quantitative Proteomics Analysis Identifies Mitochondria as Therapeutic Targets of Multidrug-Resistance in Ovarian Cancer

“…Expression levels of cofilin, trifunctional enzyme subunit alpha and isoform 1 of 3,2-trans-enoyl-CoA isomerase were found to be up-regulated in MCF-7/ADRVp cells but were unchanged in NCI_ADR/RES cells. Jiang et al investigated alterations in the mitochondrial proteome of Raji cells treated with ADR using 2D-DIGE and LTQ-MS/MS, and found altered expression of 37 proteins 33 , some of which do not agree with the findings for MCF-7/ADRVp or NCI_ADR/RES. For example, PHB, which was up-regulated in DOX-treated Raji cells, was virtually unaltered in NCI_ADR/RES cells.…”

Cancer Cell Response to Anthracyclines Effects: Mysteries of the Hidden Proteins Associated with These Drugs

“…To date, the effect of DOXO treatment on different cancer cell lines has mainly been studied by proteomic techniques [11,13,14]. To extend current observations and with the view to help translation of molecular findings toward improvements in clinical use, we focused on the effects of several clinically relevant representatives of the group of anthracycline/anthracenedione drugs.…”

“…Using a suitable protein stain, this popular and reliable technique may facilitate comprehensive quantification [10]. Several proteomic studies have been recently performed for monitoring the effect of DOXO on hepatocellular carcinoma [11], breast cancer [12], non-Hodgkin lymphoma [13], acute lymphoblastic leukemia cells [14] or the effect of DNR on pancreatic carcinoma [15] in vitro. In addition, proteomic techniques have been used for studying drug resistance mechanisms to DOXO or MTX in lung cancer cells [16,17].…”

Proteomics for identifying mechanisms and biomarkers of drug resistance in cancer

“…In a study, comparative proteomic analysis was performed in the mitochondrial proteins of adriamycin-exposed and unexposed non-Hodgkin lymphoma (NHL) Rji cells, and 37 differential proteins were identified in the adriamycintreated cells and control cells, of which heat shock protein 70 (HSP70), ATP-binding cassette transporter isoform B6 (ABCB6), and prohibitin1 (PHB1) may be related to adriamycin resistance[33]. A comparative proteomic analysis was conducted to identify the changes of mitochondrial proteins in the adriamycin-resistant human breast cancer MCF-7 cells relative to its parental MCF-7 cells.…”