Antitumorski efekat novosintetisanih kompleksa rutenijuma(II) na tumorske ćelije in vitro
Antitumor effects of newly synthesized ruthenium(II) complexes against cancer cells in vitro
Author
Čanović, PetarMentor
Zelen, Ivanka
Committee members
Mitrović, MarinaVolarević, Vladislav

Milovanović, Marija
Misirlić Denčić, Sonja
Metadata
Show full item recordAbstract
Uvod: Uprkos uspehu u lečenju različitih vrsta tumora, klinička efikasnost cisplatine
je veoma ograničena razvijanjem rezistencije tumora na lek. Kompleksi rutenijuma kao
antitumorski agensi najviše obećavaju, pokazujući citotoksičnu aktivnost kod tumora
rezistentnih na cisplatinu. Smatra se da ove osobine jedinjenja rutenijuma potiču od
njihove sposobnosti da imitiraju način vezivanja gvožđa za biomolekule kao što su
humani serum albumin i transferin. Budući da tumorske ćelije na svojim membranama
povećano eksprimiraju transferinske receptore zbog povećane potrebe za gvožđem,
dopremanje kompleksa rutenijuma do kancerskih ćelija je efikasnije u odnosu na druge
komplekse metala.
Cilj: Studija se bavi ispitivanjem potencijalne citotoksičnosti dva novosintetisana
kompleksa rutenijuma: Ru-1, [Ru(Cl-Ph-tpy)(o-pda)Cl]Cl (Cl-Ph-tpy = 4-(4'-hlorofenil)-
2,2',6',2''-terpiridin, o-pda = o-fenilendiamin) i Ru-2, [Ru(Cl-Ph-tpy)(phen)Cl]Cl(Cl-Ph-tpy = 4-
(4'-hlorofenil)-2,2',6',2''-terp...iridin,phen = 1,10-fenantrolin) na sledećim tumorskim ćelijama:
A549 – adenokarcinom pluća, MCF7 – karcinom dojke, HeLa – karcinom cerviksa, Hs 294T
– maligni melanom i na ćelijskoj liniji zdravih fibroblasta MRC-5 (kontrola).
Materijal i metod: Citotoksičnost kompleksa rutenijuma Ru-1 i Ru-2 na tumorskim i
kontrolnim, zdravim ćelijama je određivana MTT metodom. Tip ćelijske smrti kao i
relativni odnos nekrotične i apoptotične smrti tumorskih ćelija izazvane testiranim
kompleksima ispitivan je metodom protočne citometrije ćelija bojenih Annexin-om V i 7-
aminoaktinomicinom D. Faze ćelijskog ciklusa su određivane metodom bojenja
propidijum-jodidom. Aktivacija i lokalizacija ključnih proteina uključenih u proces
apoptoze je određena protočnom citometrijom bojenjem specifičnim antitelima.
Rezultati: Citotoksičnost kompleksa rutenijuma Ru-1 i Ru-2 je ispitana na četiri
tumorske ćelijske linije (A549, MCF7, HeLa i Hs294T) i na zdravim fibroblastima (MRC-
5). Naši rezultati jasno pokazuju značajno smanjenje vijabilnosti tumorskih ćelija.
Takođe, oba kompleksa rutenijuma su snažno indukovala apoptozu tretiranih ćelija sa
visokim procentom apoptotskih ćelija i zanemarljivim procentom nekrotičnih ćelija.
Zaključak: Kompleksi rutenijuma(II) pokazuju selektivnu antitumorsku aktivnost koja je,
u nekim slučajevima, i veća nego citotoksična aktivnost cisplatine. Kompleksi Ru-1 i
Ru-2 gotovo da nisu imali dejstvo na vijabilnost fibroblasta. Precizni mehanizam
delovanja kompleksa rutenijuma(II) nije u potpunosti razjašnjen. Međutim, pokazano je da
oba kompleksa smanjuju vijabilnost tumorskih ćelija indukcijom apoptoze i
zaustavljanjem ćelijskog ciklusa u određenoj fazi.
Introduction: Ruthenium complexes might be very promising candidates as antitumor agents,
showing activity in tumors which had developed resistance to cisplatin or in which cisplatin is
inactive. This is believed to be due to the ability of ruthenium to mimic iron in binding to
biomolecules, such as human serum albumin and transferrin. As cancer cells overexpress
transferrin receptors, to satisfy their increased demand for iron, ruthenium-based drugs may be
delivered more efficiently to cancer cells compared to other metal-based drugs.
Objective: The aim of our study was to investigate antitumor effect and the mechanism of action
of two newly sinthesized ruthenium complexes: Ru-1, [Ru(Cl-Ph-tpy)(o-pda)Cl]Cl (Cl-Ph-tpy = 4-
(4'-hlorofenil)-2,2',6',2''-terpiridin, o-pda = o-fenilendiamin) and Ru-2, [Ru(Cl-Phtpy)(
phen)Cl]Cl(Cl-Ph-tpy = 4-(4'-hlorofenil)-2,2',6',2''-terpiridin,phen = 1,10-fenantrolin) against
human cancer cell lines: A549 - lung cancer, MCF-7 - breast cancer, He...La - cervical cancer, Hs
294T- melanoma and one non-cancerous cell line MRC-5 - healthy fibroblasts (control).
Material and Methods: The cytotoxicity of two newly synthesized Ru(II) polypyridyl complexes
Ru-1 and Ru-2 on experimental and control group of cells was determined by the MTT assay.
Apoptosis of both control and experimental group of tumor cells was estimated by annexin Vfluorescein
isothiocyanate (FITC)/7-amino-actinomycin D (7-AAD) staining. The cell cycle
distribution was determined using propidium iodide staining. The localization and activation of
key apoptotic proteins (bax, bcl-2, cytochrome c and caspase-3) was determined by flowcytometry.
Results: The cytotoxicity of two ruthenium(II) complexes 1 and 2 was evaluated in four human
cancer cell lines (A549, MCF7, HeLa and Hs294T) and in one non-cancer cell line (MRC-5). Our
results clearly showed significant decrease of cancer cells’ viability compared to the viability of
control, non-cancerous cells, with specific individual cell lines sensitivities. Both ruthenium
complexes strongly induced apoptosis of treated cancer cells via intrinsic or mitochondrial
pathway, with high percentages of apoptotic cells and negligible percentage of necrotic cells
making these compounds suitable for further anticancer evaluation.
Conclusion: The newly synthesized ruthenium(II) complexes 1 and 2 showed selective anticancer
activity against different types of cancer cells. In some cases, their activity is even higher than that
of cisplatin in the same cells. Moreover, it is very important to delineate that these complexes had
almost no effect in tested concentrations on viability of healthy cells in vitro. The precise
mechanism of action of investigated ruthenium(II) complexes is not fully understood. However,
our results showed that both complexes decreased viability of cancer cells by induction of
apoptosis and/or by cell cycle arrest which implies their different mechanism of action on different
types of cancer cells.