Cell cycle, apoptosis, cellular uptake and whole-transcriptome microarray gene expression analysis of HeLa cells treated with a ruthenium(II)-arene complex with an isoquinoline-3-carboxylic acid ligand.
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 IC50values 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 RuT7showed 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/106cells after 6h of incubation. To gain further insight in the molecular mechanism of action of RuT7on 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 RuT7showed 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 RuT7in animal and pre-clinical studies as a potential drug candidate.
Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
Katarina K
KK
Jovanović
Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
Miljana
M
Tanić
Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, 11 000 Belgrade, Serbia.
Ivanka
I
Ivanović
Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
Nevenka
N
Gligorijević
Institute of Chemistry, Technology and Metallurgy, Centre of Chemistry, University of Belgrade, Studentski trg 12-16, Belgrade, Serbia.
Biljana P
BP
Dojčinović
Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia. Electronic address: sinisar@ncrc.ac.rs.
Siniša
S
Radulović
D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects*; D056831:Coordination Complexes / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D007546:Isoquinolines / Q000737:chemistry / Q000494:pharmacology; D012428:Ruthenium / Q000737:chemistry / Q000494:pharmacology; D059467:Transcriptome / Q000187:drug effects*
D056831:Coordination Complexes; D007546:Isoquinolines; C000606691:isoquinoline-3-carboxylic acid; D012428:Ruthenium
Anticancer drugs; Apoptosis; HeLa; Microarray; Ruthenium(II)