Formation of multicellular tumor spheroids induced by cyclic RGD-peptides and use for anticancer drug testing in vitro.
Development of novel anticancer formulations is a priority challenge in biomedicine. However, in vitro models based on monolayer cultures (2D) which are currently used for cytotoxicity tests leave much to be desired. More and more attention is focusing on 3D in vitro systems which can better mimic solid tumors. The aim of the study was to develop a novel one-step highly reproducible technique for multicellular tumor spheroid (MTS) formation using synthetic cyclic RGD-peptides, and to demonstrate availability of the spheroids as 3D in vitro model for antitumor drug testing. Cell self-assembly effect induced by addition of both linear and cyclic RGD-peptides directly to monolayer cultures was studied for 12 cell lines of various origins, including tumor cells (e.i. U-87 MG, MCF-7, M-3, HCT-116) and normal cells, in particular L-929, BNL.CL2, HepG2. Cyclo-RGDfK and its modification with triphenylphosphonium cation (TPP), namely cyclo-RGDfK(TPP) in a range of 10-100μM were found to induce spheroid formation. The obtained spheroids were unimodal with mean sizes in a range of 60-120μm depending on cell line and serum content in culture medium. The spheroids were used as 3D in vitro model, in order to evaluate cytotoxicity effects of antitumor drugs (doxorubicin, curcumin, temozolomide). The developed technique could be proposed as a promising tool for in vitro test of novel antitumor drugs.
Polymers for Biology Laboratory, Shemyakin-Ovchinnikov Institute of Bioorganic chemistry of the Russian Academy of Sciences, 117997, Miklukho-Maklaya 16/10, Moscow, Russia; CNRS UMR 7199, University of Strasbourg, Laboratoire de Conception et Application de Molécules Bioactives, 74 route du Rhin, 67401 Illkirch Cedex, France. Electronic address: roman.akasov@gmail.com.
Roman
R
Akasov
Polymers for Biology Laboratory, Shemyakin-Ovchinnikov Institute of Bioorganic chemistry of the Russian Academy of Sciences, 117997, Miklukho-Maklaya 16/10, Moscow, Russia.
Daria
D
Zaytseva-Zotova
Synthesis of Peptides and Polymer Microspheres Laboratory, Institute of Macromolecular Compounds of the Russian Academy of Sciences, 199004, Bolshoy pr. 31, Saint-Petersburg, Russia.
Sergey
S
Burov
Synthesis of Peptides and Polymer Microspheres Laboratory, Institute of Macromolecular Compounds of the Russian Academy of Sciences, 199004, Bolshoy pr. 31, Saint-Petersburg, Russia.
Maria
M
Leko
CNRS UMR 7213, University of Strasbourg, Laboratoire de Biophotonique et Pharmacologie, 74 route du Rhin, 67401 Illkirch Cedex, France.
Monique
M
Dontenwill
CNRS UMR 7199, University of Strasbourg, Laboratoire de Conception et Application de Molécules Bioactives, 74 route du Rhin, 67401 Illkirch Cedex, France.
Manuela
M
Chiper
CNRS UMR 7199, University of Strasbourg, Laboratoire de Conception et Application de Molécules Bioactives, 74 route du Rhin, 67401 Illkirch Cedex, France.
Thierry
T
Vandamme
Polymers for Biology Laboratory, Shemyakin-Ovchinnikov Institute of Bioorganic chemistry of the Russian Academy of Sciences, 117997, Miklukho-Maklaya 16/10, Moscow, Russia.
Elena
E
Markvicheva
D000970:Antineoplastic Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical / Q000379:methods; D003474:Curcumin / Q000494:pharmacology; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology; D004317:Doxorubicin / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D045325:HCT116 Cells; D056945:Hep G2 Cells; D006801:Humans; D061986:MCF-7 Cells; D009842:Oligopeptides / Q000494:pharmacology*; D010456:Peptides, Cyclic / Q000494:pharmacology*; D018874:Spheroids, Cellular / Q000187:drug effects*; D000077204:Temozolomide
D000970:Antineoplastic Agents; D009842:Oligopeptides; D010456:Peptides, Cyclic; C047981:arginyl-glycyl-aspartic acid; D003606:Dacarbazine; D004317:Doxorubicin; D003474:Curcumin; D000077204:Temozolomide
Cell self-assembly; Curcumin (PubChem CID: 969516); Cyclic RGD-peptides; Cyclo-RGDfK (PubChem CID: 10196873); Cytotoxicity; Doxorobicin (PubChem CID: 443939); Drug screening; Multicellular tumor spheroids; Temozolomide (PubChem CID: 5394)