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Volum 08 Numarul 4, 2002

219 Editorial

The Annual Conference of Romanian Society for Radiotherapy and Oncology (RSRO) who took place between 9-12 of October 2002 had as one of the main research subject Experimental models and radiochemotherapy basic principles. Our review is dedicated to most important papers exposed at the Conference. All the papers are the result of the scientific work in Radiation Oncology Research Center RORC. This center is a join venture between the Department of Radiotherapy of the University of Medicine and Pharmacy Cluj, Cancer Institute Cluj, and Romanian Society of Radiation Oncology. One year before in another number of our review (4/200 I) V.Cernea at al. have presented the RORC which at the time was in full organisation and the first equipements arrived. At just a few months from its inauguration in june 2002 is a honor for all research team members to can present their work in some new laboratories even for Romania, as well as some general papers.

The first paper published by V. Cernea at al., some in vitro and in vivo experimental models used in radiobiology are presented in order to test citostatic drugs and their practical approach in molecular biology study. The economic and technical advantages of cell cultures and multicelular spheroides are also presented for radio-chemotherapy combination effects. The main subject is the description of experimental models on inbred and nude mice to explain some fundamental aspects of cancerogenesis processes in order to test some drugs and types and regimens for radiation fractioning.

The cellular survive curves semnification and the most used models in appreciating the radiation effects at cellular level is presented by Ioana Brie. Far from being only theories, these models elaboration of cellular response to radiation permit the correlation of experimental results obtained in laboratory with clinical results in order to ameliorate the treatments. Earn a special accent on the first radiobiology experiments on cell culture in the laboratory with the same name of RORC, first time tried in our country.

  1. GhiIezan presents in his paper important correlation between cellular signaling circuits and their manipulation in therapeutic application. The selective inhibition of proliferation and angiogenesis signals at tumor level by using monoclonal antibodies anti extracellular domain of growth factor receptors can sustain a therapeutic argument. The general structure of cellular signaling circuits is presented as well as the first clinical results of some new therapeutic modalities to inhibit or stop some of those which manifest a sinergic effect with radiotherapy.

Luminita Lelutiu offers an interesting incursion in molecular citogenetics and its major goal in clinical oncology. The value of new and specific techniques like FISH and spectral cariptyping are presented as highly precise diagnostic modality in citogenetics.

The DNA analize through nanotechnology is presented by O. Bălăcescu paper. One of the most spearhead equipment in RORC is a bioalnalyzor produced by Agilent Technologies which give the possibility to the researcher to made the quality control for tumor DNA used in PCR technique. The chip technology for nucleic acids is really revolutionary in molecular biology for its accuracy and small quantities of genetic material.

loana Berindan Neagoe presents the first reserach in Molecular Biology Laboratory for p53 gene in breast cancer, a study made in Oncological Institute in order to obtain the mutation specific for this gene. Some specific techniques in molecular biology are presented like DNA extraction ,PCR, and automated sequencing.

Doina Daicoviciu at al. present the matrix metalloproteases significance in brest tumors. So the gelatinase A level is highly expressed in mammary carcinomas in comparison with benign tumors of the breast.

The immunofenotype analyze in differential diagnostic ofcronics lymph proliferation is underlined in article presented by Olga Soriţău at al. The accuracy of diagnostic can be improved by tlowcytometry.

The short communication of Ioana Berindan Neagoe is on cell adhesion molecules in invasion and metastasis. News in Oncology inform us on three imporant events. The UICC Congress in Oslo in 30 of June 5 of July is presented by Pertonela Rusu, with accent on lung cancer. The phiysics course for clinical radiotherapyin Leuven (25-29 of August) is presented by A. Veres and G. Kacso inform us on 21-st ESTRO Conference at Prague between 17 to 21 september.

  1. Cernea, Ioana Berindan Neagoe

221 Model and Tumor Systems in Radio-Chemotherapy

  1. Cernea 1,2, N. Ghilezan 1,2, Ioana Berindan Neagoe2, O. Balacescu2, Rodica Rişcă3

1UMPh „Iuliu Haţieganu” Cluj-Napoca

2Cancer Institute „Prof I. Chiricuţă

3BIONA Cluj-Napoca

Purpose: To present the tumor systems most used in the study or radio-chemotherapy and in more general sense in tumor biology. Definitions: Cell cultures are experimental in vitro systems, which allow the study of radiation and chemotherapy effects at cellular level. They are most used system today because of low cost and experiments reproducibility. Their limits are the lack of cell-to-cell interactions, and the difficulty to extrapolate the results to human tumors. Multicellular tumor spheroids are three-dimensional structures composed of tumor cells. Because of their particular architectural characteris- tics, are demonstrated to be extremely useful in testing radio therapeutic protocols, including dose rate and fractionation and the effects of combined radio-chemotherapy treatments. Xenografts are human tumors transplanted at immunologicaly- compromised animals. The tumor parenchyma can be investigated in conditions similar to human tumors, but the studies concerning the vasculature are much less extrapolable to human tumors because of the stroma, which is of murine origin. Conclusions: Model and tumor systems are part of cancer research and have determined considerable progress in tumor biology and treatment.

Key words: models, cell cultures, spheroids, xenografts, radio-chemotherapy.

Radioterapie & Oncologie Medicală, 2002, 4:221-228

229 Cell Survival Curves – Tools for Estimating Radiation Effects

Ioana Brie2, V. Cernea1,2, Maria Perde1, Olga Soriţău1, Eva Szikszai1, Piroska Virag1, I.D. Postescu1, N.Ghilezan1,2

1Cancer Institute „Prof I.Chiricuţă” Cluj-Napoca

2UMPh „Iuliu Haţieganu” Cluj-Napoca

Research in experimental radiobiology covers studies at the cell, animal and human levels, dealing with the nature of radiation damage. It operates with models in order to build up theories of the radiation action both in vitro and in vivo, as well to core late the experimental studies with clinical results, all these to improve therapy. Cell-survival curves are plots of survival fraction versus dose. Among the models proposed to describe their shape, the linear-quadratic one has important clinical applications. The survival curves can be obtained in vitro, in vivo and also in spheroid cultures. By comparing the survival curves of cells before and after malignant transformation, no difference in intrinsic radiosensitivity between normal and malignant cells of the same histological type was found. The differences in radiation responses are due to other factors, such as hypoxia. The survival curves are useful tools to compare two or more cellular populations, to estimate the radiation dose which can sterilize a tumor cell population and to study the factors that modify the survival of irradiated cells.

Key words: radiobiology, survival curves, models, radiosensitivity

Radioterapie & Oncologie Medicală, 2002, 4:229-235

236 Biological Modifiers of the Radiation Response: the Inhibition of the Cellular Signaling Pathways

N.Ghilezan1,2

1UMPh „Iuliu Haţieganu” Cluj-Napoca

2Cancer Institute „Prof I. Chiricuţa” Cluj-Napoca

The signaling pathways for tumor cells are characterized by the overexpression of the receptors for growth factors which are strong signal generators for proliferation, survival and angiogenesis, in contrast with the normal cells that are strongly controlled. The selective inhibition of these pathways is nowadays possible through monoclonal antibodies directed to the extramembranal part of the receptors or by small molecules tyrozine kinase inhibitors that operate inside the cell. The general principles of the signaling pathways are presented as well the first clinical results of this new therapeutic approach

which promises a great synergistic efficacy in combination with radiation therapy.

Key words: anti-EGFR monoclonal antibodies, small molecules tyrozin-kinaze inhibitors.

Radioterapie & Oncologie Medicaid, 2002, 4:236-239

240 The Contribution of Molecular Cytogenetics at the Progress of Clinical Oncology

Luminiţa Leluţiu1, C.D. Olinici1,2

1Dept. of Pathology, UMPh „Iuliu Haţieganu” Cluj-Napoca

2Cancer Institute „Prof I. Chiricuţă” Cluj-Napoca

In the late 50s, the era of clinical cytogenetics began with technical developments which significantly improved tissue culture and chromosome preparation. Starting in the late 60s with the description of Q-banding, the essential techniques developed during the early alld mid 70s are still applied as routine diagnostic techniques at the present time. In the last few years the introduction of molecular-cytogenetics proved soon its value in supplementing the routine approaches. While fluorescence in situ hybridisation (FISH) has become a standard technique, new ones such as comparative genomic hybridisation (CGH), spectral karyotyping (SKY) and multi-color FISH(M-FISH) which rely basically on in situ hybridisation, have shown their potential for diagnostic purposes. Thus, it is obvious that for comprehensive diagnostic cytogenetics a combination of both traditional and new cytogenetic techniques are required in a number of case.

Keywords: cytogenetics, chromosome, karyotype, molecular biology.

Radioterapie & Oncologie Medicală. 2002. 4:240-243

244 Detection and Quantification of Tumor – DNA Through the Use of Microfluidic Technologies

O.Bălăcescu1, Ioana Berindan Neagoe1, N.Ghilezan1,2, V.Cernea1,2, Liana Cosma2

1Cancer Institute „Prof I.Chiricuţă”, Cluj-Napoca

2UMPh „Iuliu Haţieganu” Cluj-Napoca

The new era of nanotechnology was build in ‘he last decade as a consequence of translational research from laboratory research to clinic. By using lab machines like Bioanalizor 21 00 which achieve the market last year, the Research Centre of Radiation Oncology has the possibility to use for DNA, RNA and protein analysis this new tool in Molecular Biology in order to obtain quality control for tumour manipulation. The molecular biology Department use the nanochips for PCR amplification fragments, RNA quality as a consequence of nucleic acids extraction, amplification or fragments. Our results demonstrates the high difference between usual gels used in laboratory and accuracy and reproducibility of nanochips which have also the advantage of using unsignificant quantities of sample. Through the use of microfluidic technology, only 1 μl of nucleic acid sample is required for separation in micro-channels that are filled with a sieving polymer and a fluorescent dye. The advantages of bioanalyzer boldly stand out with respect to sensitivity, linear dynamic range and sizing capability over a broad range of fragment sizes. The attractive cost and performance of the nanotechnology make from this new methods one of the future biotechnologies instrument.

Key words: nanotechnology, chip, biotechnology, DNA, RNA.

Radioterapie & Oncologie Medicală. 2002, 4:244-248

249 Molecular Biology Techniques for Optimization of Polymerase Chain Reaction in Order to Detect the p53 Gene Mutations in Breast Carcinomas

Ioana Berindan-Neagoe1, O. Bălăcescu1, Liana Cosma1, N.Ghilezan1,2, V.Cernea1,2, Andreea Şodolescu2

1Cancer Institute „Prof Dr. I. Chiricuţă” Cluj-Napoca

2UMPh „Iuliu Haţieganu” Cluj-Napoca

One of the first characterised genes, demonstrating high implications in prognostic and malignant tumoe evolution is the p53 gene, which remain a major stone for new era of biotechnology in cancer study. One of the major goals of the molecular biology of cancer is to identify new factors and pathways for cancer therapy, more efficient and convenient for the patient. Our study tried to optimise one of the most important methods in molecular biology, the polymerase chain reaction (PCR) in order to establish the exons 4, 5, 6, 7,8, and 9 for future sequencing. We extracted and amplified the DNA from 25 patients treated with surgery between 2001-2002. We established the methods for extraction, purification and quantitative evaluation of nucleic acids, done for the first time in RCRO, to prepare the amplified exons foe sequence detection.

Key words: DNA extraction, polymerase chain reaction, optimisation, p53 gene, exons.

Radioterapie & Oncologie Medicală. 2002. 4:249-254

255 Study on Proteolytic Activity in Breast Tumor Tissue

Doina Daicoviciu, I.D. Postescu, Daniela Grecea, Corina Tatomir, Angela Răşinariu, Gabriela Cherecheş

Cancer Institute „Prof Dr: l Chiricuţă”

Matrix metalloproteinases (MMP) playa major role in degrading extracellular matrix (MEC) during invasion and metastasis processes. Gelatinase A (MMP-2) and gelatinase B(MMP-9) are proteolytic enzymes hydrolyzing type IV collagen from MEC. The aim of the study was to compare the activity of geIatinase A in breast tumor tissues from early breast cancer patients with the activity in tissues obtained from patients with benign diseases. In breast tumors two bands of gelatinolytic activity were observed: one corresponding to 72kDa pro-enzyme, the other corresponding to 68kDa mature enzyme. The level of gelatinase A total activity was higher in tumor tissues vs. benign breast diseases tissues. No significant correlations between gelatinase A activity and clinical or histological characteristics of the patients were observed.

Key words: gelatinase A, breast tumor tissues, zymography.

Radioterapie & Oncologie Medicală, 2002, 4:255-260

261 Immunophenotyping Contribution to Differential Diagnosis in Chronic Lymphoproliferative Diseases

Olga Soriţău, Mariana Paţiu, Piroska Virag, Eva Fischer, Carmen Ştugren

Cancer Institute Prof Dr: I. Chiricuţă”

The purpose of this paper is to establish the limits of fluorescence microscopy and a minimal monoclonal antibody panel used in differential diagnosis of chronic Iymphoproliferations. These hematological diseases were studied on 16 patients from Hematology Department. Evidence of cellular clonality was possible in 8 cases with B Iymphoproliferation and 3 cases with T Iymphoproliferation. The conclusions are: 1. accurate diagnosis requires careful correlation of histologic, hematologic, immunologic and clinical findings; 2. the necessity of a flow cytometric analyse system and a more large monoclonal antibodies panel.

Key words: chronic Iymphoproliferative disease, immunophenotyping.

Radioterapie & Oncologie Medicală. 2002. 4:261-264

266 Advances in Lung Cancer in the Light of the 18th UICC International Cancer Congress 30 June 5 July, 2002, Oslo, Norway

Petronela Rusu

270 Basic Physics for Clinical Radiotherapy. August 25-29, 2002, Leuven, Belgium

  1. Veres

271 Report on the 21th ESTROAnnual Meeting, September 17-21,2002, Prague, Czech Republic (I)

G.Kacso

274 Calcium Dependent Cell Adhesion Molecules. Cadherins

Ioana Berindan-Neagoe

277 CONTENTS OF Journal of Romanian Society of Radiation Therapyan Medical Oncology: Volumul VIII, 2002

281 TOPIC’S INDEX 2002

284 AUTHOR’S INDEX 2002

285 MEMBERS OF ROMANIAN SOCIETY OF RADIATION THERAPY AND MEDICAL ONCOLOGY 2002

286 GUIDELINES FOR AUTHORS

288 ERRATUM