Identification Of Predictive In Vitro Molecular Markers Of The Response To The Treatment To Oxaliplatin In Colorectal Cancers. Utilization On In Vivo Models In Translational Research

Host institute of the project: “I. Chiricuta” Cancer Institute – Comprehensive Cancer Center, Cluj-Napoca, Republicii str., nr. 34-36, tel/fax: 0264-598361/0264-439260
Project coordinator: Piroska Virag – PhD, principal biologist
Internal collaborators: Ioana Neagoe – PhD, principal chemist; Ovidiu Balacescu – PhD, principal chemist; Eva Fischer-Fodor - principal chemist, young researcher; Claudia Burz – MD, young reseracher; Loredana Balacescu – physicist, young researcher.
Project enrollment period: 2009-2011
Budjet: 1 000 000 lei
Financing: National Council of Scientific Research in Higher Education (NURC)
Time schedule: Year 2009:
Phase I.
1. Purchasing the media and reagents necessary for the analyses
1.1. Purchasing the resources necessary for the colorectal cancer cell lines (cell culture media, supplies)
1.2. Purchasing cell cycle evaluation kit
2. In vitro study on Oxaliplatin sensitive and resistant cell lines
2.1. Development of an Oxaliplatin resistant cell line by treating sensitive cells with risen concentrations of Oxaliplatin
2.2. Establishment of the maximum lethal dose of Oxaliplatin with MTT test; study of the influence of the administered doses

Phase II.
1. Analyses of the cell cycle modifications and apoptosis induced by Oxaliplatin on sensitive and resistant cell lines
1.1. Evaluation of the cell cycle modifications and apoptosis induced by Oxaliplatin, on sensitive and resistant cell lines
2. Preparation of functional genomics study by microarray and RT-PCR
2.1. Purchasing reagents necessary to microarray and RT-PCR techniques

Obtained results:
Phase I.
1. Theoretical and practical preparation of the study by meetings and round tables; presentation of the grant, the current status of the colorectal cancer in Romania and of the study methods. Purchasing specific materials required for the cultivation of Colo320 colorectal cell line: cell culture media, supplements and cell viability reagent (MTT-dimethylthiazol tetrazolium bromide). Purchasing cell cycle evaluation kit, prepearing the next phase of the project.
2. The colorectal cell line Colo320 was treated with risen concentrations of Oxaliplatin, obtaining an Oxaliplatin resistant cell line. We performed MTT cell viability test in order to evaluate the influence of the administered doses on the cell survival.

Phase II.
1. Oxaliplatin induced cell cycle modifications and apoptosis were evaluated, on sensitive and resistant cell lines.
2. In order to prepare the functional genomics study we purchased reagents necessary to microarray and RT-PCR tests.