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Synthesis and Evaluation of a Ruthenium-based Mitochondrial Calcium Uptake Inhibitor
Published on: October 26, 2017
Cell cycle, apoptosis, cellular uptake and whole-transcriptome microarray gene expression analysis of HeLa cells
Katarina K Jovanović1, Miljana Tanić1, Ivanka Ivanović2
1Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
Abstract:
Ruthenium(II)-arene complexes are promising drug candidates for the therapy of solid tumors. In previous work, seven new compounds of the general formula [Ru(η6-p-cymene)(L1-7)Cl] were synthesized and characterized, of which the complex with L=isoquinoline-3-carboxylic acid (RuT7) was two times as active on HeLa cells compared to normal cell line MRC-5, as indicated by IC50 values determined after 48h of incubation (45.4±3.0 vs. 84.2±5.7μM, respectively). In the present study, cell cycle analysis of HeLa cells treated with RuT7 showed S phase arrest and an increase in sub-G1 population. The apoptotic potential of the title compound was confirmed with the Annexin V-FITC/PI assay together with a morphological evaluation of cells using fluorescent microscopy. Analysis of the intracellular accumulation of ruthenium showed 8.9ng Ru/106 cells after 6h of incubation. To gain further insight in the molecular mechanism of action of RuT7 on HeLa cells, a whole-transcriptome microarray gene expression analysis was performed. Analysis of functional categories and signaling and biochemical pathways associated with the response of HeLa cells to treatment with RuT7 showed that it leads the cells through the intrinsic (mitochondrial) apoptotic pathway, via indirect DNA damage due to the action of reactive oxygen species, and through direct DNA binding of RuT7. Statistical analysis for enrichment of gene sets associated with known drug-induced toxicities identified fewer associated toxicity profiles in RuT7-treated cells compared to cisplatin treatment. Altogether these results provide the basis for further development of RuT7 in animal and pre-clinical studies as a potential drug candidate.
Insights
Ruthenium(II)-arene complex RuT7 shows significant anti-cancer activity against HeLa cells by inducing apoptosis and cell cycle arrest. This promising compound exhibits lower toxicity compared to cisplatin, warranting further pre-clinical development.
Area of Science:
- Medicinal Chemistry
- Cancer Therapy
- Ruthenium Complexes
Background:
- Ruthenium(II)-arene complexes are investigated as potential anti-cancer agents.
- Previous studies synthesized seven novel compounds, including RuT7 with isoquinoline-3-carboxylic acid.
- RuT7 demonstrated enhanced activity against HeLa cancer cells compared to normal cells.
Purpose of the Study:
- To elucidate the mechanism of action of RuT7 in HeLa cells.
- To evaluate the apoptotic potential and cell cycle effects of RuT7.
- To compare the toxicity profile of RuT7 with cisplatin.
Main Methods:
- Cell cycle analysis (flow cytometry) and Annexin V-FITC/PI assay.
- Intracellular ruthenium accumulation measurement.
- Whole-transcriptome microarray gene expression analysis.
Main Results:
- RuT7 induced S phase arrest and increased sub-G1 population in HeLa cells.
- Apoptosis was confirmed through Annexin V-FITC/PI assay and morphological studies.
- Gene expression analysis revealed RuT7 activates the intrinsic apoptotic pathway via ROS and direct DNA binding, with a favorable toxicity profile versus cisplatin.
Conclusions:
- RuT7 exhibits potent anti-cancer effects through apoptosis induction and cell cycle disruption.
- The compound's mechanism involves ROS-mediated DNA damage and direct DNA binding.
- RuT7 presents a promising therapeutic candidate with reduced toxicity for further pre-clinical evaluation.
