Protein Expression Profile of HT-29 Human Colon Cancer Cells after Treatment with a Cytotoxic Daunorubicin-GnRH-III Derivative Bioconjugate

Abstract

Targeted delivery of chemotherapeutic agents is a new approach for the treatment of cancer, which provides increased selectivity and decreased systemic toxicity. We have recently developed a promising drug delivery system, in which the anticancer drug daunorubicin (Dau) was attached via oxime bond to a gonadotropin-releasing hormone-III (GnRH-III) derivative used as a targeting moiety (Glp-His-Trp-Lys(Ac)-His-Asp-Trp-Lys(Dau = Aoa)-Pro-Gly-NH2; Glp = pyroglutamic acid, Ac = acetyl; Aoa = aminooxyacetyl). This bioconjugate exerted in vitro cytostatic/cytotoxic effect on human breast, prostate and colon cancer cells, as well as significant in vivo tumor growth inhibitory effect on colon carcinoma bearing mice. In our previous studies, H-Lys(Dau = Aoa)-OH was identified as the smallest metabolite produced in the presence of rat liver lysosomal homogenate, which was able to bind to DNA in vitro. To get a deeper insight into the mechanism of action of the bioconjugate, changes in the protein expression profile of HT-29 human colon cancer cells after treatment with the bioconjugate or free daunorubicin were investigated by mass spectrometry-based proteomics. Our results indicate that several metabolism-related proteins, molecular chaperons and proteins involved in signaling are differently expressed after targeted chemotherapeutic treatment, leading to the conclusion that the bioconjugate exerts its cytotoxic action by interfering with multiple intracellular processes.

Mass spectrometric characterization of the zein protein composition in maize flour extracts upon protein separation by SDS‐PAGE and 2D gel electrophoresis

“…The resolution is high enough to distinguish a particular product from another yet related product . The fact that this combination of modern analytical methods applied here was successful for characterizing rather complex mixtures of closely related natural proteins, it had been anticipated from results of studies which had applied these methods for the characterization of plant material as well as of studies with human blood and tissues samples . However, one had to consider that the genomic information in the databases reflects the DNA sequence of a model plant and that differences in coding sequences within varieties exist.…”

Synthesis, characterization and systematic comparison of FITC‐labelled GnRH‐I, ‐II and ‐III analogues on various tumour cells

“…In experimental setups, GnRH‐III significantly suppressed the proliferation of human cancer cells, which overexpress GnRH‐R . Therefore, also daunorubicin‐containing GnRH‐III conjugates were synthesized and investigated in vitro and in vivo . These conjugates were well tolerated and had a significant antitumour effect on colon carcinoma‐bearing mice .…”

Hesperitin enhances the ability of daunorubicin by co-loading with MPEG-PCL nanoparticles to induce apoptosis in gastric cancer

“…Hesperitin is reported to possess anti-cancer ability, such as prostate cancer and esophageal cancer [22,36,37]. Unfortunately, its efficiency was limited due to high hydrophobicity [20,25,47] [26,27,38,39]. Daunorubicin is accompanied with cardiac toxicity, which restrained the dosage that could be used [40].…”