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

Abstract

A comprehensive proteome map of T-lymphoblastic leukemia cells and its alterations after daunorubicin, doxorubicin and mitoxantrone treatments was monitored and evaluated either by paired comparison with relevant untreated control and using multivariate classification of treated and untreated samples. With the main focus on early time intervals when the influence of apoptosis is minimized, we found significantly different levels of proteins, which corresponded to 1%–2% of the total amount of protein spots detected. According to Gene Ontology classification of biological processes, the highest representation of identified proteins for all three drugs belong to metabolic processes of proteins and nucleic acids and cellular processes, mainly cytoskeleton organisation and ubiquitin-proteasome pathway. Importantly, we observed significant proportion of changes in proteins involved in the generation of precursor metabolites and energy typical for daunorubicin, transport proteins participating in response to doxorubicin and a group of proteins of immune system characterising response to mitoxantrone. Both a paired comparison and the multivariate evaluation of quantitative data revealed daunorubicin as a distinct member of the group of anthracycline/anthracenedione drugs. A combination of identified drug specific protein changes, which may help to explain anti-cancer activity, together with the benefit of blocking activation of adaptive cancer pathways, presents important approaches to improving treatment outcomes in cancer.

Reticulocytes Are an Enriched Source of the RUVBL1 Protein

“…The RUVBL1 and GAPDH antibodies used each detected single bands in the human RBC samples at ∼50 and ∼37 kDa, respectively, on Western blots, which is in accordance with what has previously been reported in the literature [7,9,11] and by various manufacturers.…”

“…After removal of the supernatant, the RBCs were boiled in a reducing buffer (10% glycerol, 2 g% SDS, 1% 1 M Tris-HCl [pH 6.8], 5% 2-mercaptoethanol, 0.005 g% bromophenol blue), electrophoresed by SDS/10% PAGE, and transferred to PVDF membranes (Bio-Rad Laboratories). These were blocked in 3% nonfat dry milk in Tris-buffered saline (TBS) with 0.05% Tween 20, then incubated with a validated affinity-isolated Prestige Antibody to RUVBL1 from rabbit (#HPA019947, Sigma-Aldrich, developed and validated by the Human Protein Atlas project and also tested for Western blotting [9]) as well as anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH [D16H11] XP® Rabbit mAb; Cell Signaling). The blots were then washed, incubated with anti-rabbit IgG (Fc) alkaline phosphatase conjugate (Promega), and washed again.…”

Polyamine analog TBP inhibits proliferation of human K562 chronic myelogenous leukemia cells by induced apoptosis

“…Doxorubicin is one of the first-line drugs in the clinical treatment of chronic myeloid leukemia, but long-term use of this drug can lead to drug resistance and reduce its therapeutic effect ( 15 , 16 ). One clinical strategy to reduce the drug resistance of tumor cells and enhance the antitumor effect is combination therapy using two or more drugs.…”

Estrogen Enhances the Expression of the Multidrug Transporter Gene ABCG2—Increasing Drug Resistance of Breast Cancer Cells through Estrogen Receptors

“…In the MCF-7/MX–resistant cell line, the up-regulated ABCG2 requires ATP to support the major mechanism of drug resistance, and mitochondria are mainly responsible for providing cellular energy, and MX cytotoxicity may affect the mitochondrial inner membrane proteins [ 30 ] and the mitochondrial respiratory chain [ 31 ]. We, hence, investigated mitochondrial-related gene expression and cell functions further.…”

“…The cDNA was then used for PCR amplification. The primer sets for PCR amplification were as follows: ABCG2 (product size 429 bps) [ 30 ] forward 5′-TTATCCGTGGTGTGTCTGGA-3′, reverse 5′-CCTGCTTGGAAGGCTCTATG-3′; ATP synthase α subunit (product size 171 bps) forward 5′-TTGTGGTGTCGGCTACGG-3′, reverse 5′-CGGCGGAGCAACAGAGA-3′; COX VIc (product size 317 bps) [ 23 ] forward 5′-GAAGGACGTTGGTGTTGAGG-3′, reverse 5′-CTAGGGAATTCAACCTGAAG-3′. β-actin (product size 644 bps) [ 39 ] forward 5′-50-ACGTTATGGATGATGATATCG-3′, reverse 5′-CTTAATGTCACGCACGATTTC-3′.…”