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Scripta Scientifica Medica

Application and optimization of a complex approach for identification of multi-target anticancer agents

T. Zhivkova, D. Dinev, L. Dyakova, M. Georgieva, G. Miloshev, G. Marinescu, G. Luchev, I. Pashapur, D. C. Culita, L. Patron, R. Alexandrova

Abstract

Drug resistance and toxic side effects are among the main obstacles preventing successful treatment of neoplastic diseases. Development and application of agents that have the capability to interact selectively with two or more macromolecular targets is a promising strategy to overcome these challenges. The identification of compounds with anticancer potential requires a complex approach based on a wide spectrum of model systems and methods with different molecular/cellular targets and mechanisms of action. The optimization and introduction of such an approach in our laboratory was the aim of the study presented.

The cytotoxic activity of various synthetic compounds with different structure and physico-chemical character­istics (cisplatin, disulfiram, Cu(II) complexes with Schiff bases, etc.) was evaluated in short-term (24-72h, with monolayer cultures) and long-term (30-45 days, with 3D cancer cell colonies) experiments by the MTT test, neu­tral red uptake cytotoxicity assay, crystal violet staining, trypan blue dye exclusion technique, lactate dehydro­genase activity assay, double staining with acridine orange and propidium iodide, Annexin V/FITC assay, Com­et assay and colony-forming method. The investigations were performed using permanent cell lines established from chicken (hepatoma), rat (sarcoma) and human (breast cancer, cervical carcinoma, liver cancer, non-small cell lung cancer, glioblastoma multiforme) cancers of different etiology, histological type and expression of hor­monal receptors (in the case of breast cancer).

The conditions for carrying out these methods were optimized and their advantages and disadvantages were specified. Information regarding the influence of the compounds investigated on cell survival and proliferation was obtained. Our results indicate that in order to obtain adequate experimental data, the approach described above has to be adapted to each individual `cell line - compound` system.

Acknowledgements: This study was supported by Grant -DCOST 01/16 from 17.08.2017, COST Action CA15135 and a bilateral project between the Bulgarian Academy of Sciences and the Romanian Academy.





DOI: http://dx.doi.org/10.14748/ssm.v49i0.4837

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About The Authors

T. Zhivkova
IEMPAM
Bulgaria

D. Dinev
IEMPAM
Bulgaria

L. Dyakova
IEMPAM
Bulgaria

M. Georgieva
IEMPAM
Bulgaria

G. Miloshev
IEMPAM
Bulgaria

G. Marinescu
IEMPAM
Bulgaria

G. Luchev
IEMPAM
Bulgaria

I. Pashapur
IEMPAM
Bulgaria

D. C. Culita
IEMPAM
Bulgaria

L. Patron
IEMPAM
Bulgaria

R. Alexandrova
IEMPAM
Bulgaria

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