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Response Profiling Using Shotgun Proteomics Enables Global Metallodrug Mechanisms of Action To Be Established
Dominique Kreutz1, Andrea Bileck1, Kerstin Plessl2
1Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna, Waehringer Strasse 38, 1090, Vienna, Austria.
Abstract:
Response profiling using shotgun proteomics for establishing global metallodrug mechanisms of action in two colon carcinoma cell lines, HCT116 and SW480, has been applied and evaluated with the clinically approved arsenic trioxide. Surprisingly, the complete established mechanism of action of arsenic trioxide was observed by protein regulations in SW480, but not HCT116 cells. Comparing the basal protein expression in the two cell lines revealed an 80 % convergence of protein identification, but with significant expression differences, which in turn seem to affect the extent of protein regulation. A clear-cut redox response was observed in SW480 cells upon treatment with arsenic, but hardly in HCT116 cells. Response profiling was then used to investigate four anti-cancer metallodrugs (KP46, KP772, KP1339 and KP1537). Proteome alterations were mapped to selected functional groups, including DNA repair, endocytosis, protection from oxidative stress, protection from endoplasmatic reticulum (ER) stress, cell adhesion and mitochondrial function. The present data suggest that knowledge of the mechanism of action of anti-cancer metallodrugs and improved patient stratification strategies are imperative for the design of clinical studies.
Insights
Shotgun proteomics revealed differential metallodrug responses in colon cancer cells. Arsenic trioxide’s mechanism was clear in SW480 cells but not HCT116, highlighting cell-specific protein regulation impacts.
Area of Science:
- Proteomics and Cancer Biology
- Metallodrug Mechanism of Action Studies
Background:
- Understanding metallodrug mechanisms is crucial for cancer therapy.
- Colon carcinoma cell lines (HCT116, SW480) exhibit distinct basal protein expression profiles.
- Shotgun proteomics offers a global approach to analyze drug-induced proteome alterations.
Purpose of the Study:
- To establish metallodrug mechanisms of action using response profiling.
- To compare drug responses in HCT116 and SW480 colon cancer cell lines.
- To investigate the impact of basal protein expression on drug-induced regulatory effects.
Main Methods:
- Shotgun proteomics was employed for response profiling.
- Arsenic trioxide was used as a benchmark metallodrug.
- Four additional anti-cancer metallodrugs (KP46, KP772, KP1339, KP1537) were investigated.
Main Results:
- Arsenic trioxide's mechanism was fully elucidated in SW480 cells, but not HCT116 cells, due to differential protein regulation.
- Significant differences in basal protein expression between cell lines affected drug response extent.
- A distinct redox response was observed in SW480 cells upon arsenic treatment.
- Proteome alterations mapped to functional groups including DNA repair, endocytosis, and stress responses.
Conclusions:
- Cell-specific protein expression significantly influences metallodrug efficacy and mechanism elucidation.
- Response profiling is a valuable tool for dissecting metallodrug actions.
- Further research is needed to optimize patient stratification for clinical metallodrug studies.
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