The probability to develop lung cancer before 80 years of age is 1.67 and 0.18% for the male and female populations of Kharkov, respectively; the probability to develop large-intestine cancer is 0.92 and 0.49%, respectively. The correlation coefficient (r) of the age of manifestation (AM) of cancer in parent-offspring pairs is 0.47. These correlation coefficients for the father-son, mother-daughter, mother-son, and father-daughter pairs are 0.64, 0.49, 0.44, and 0.37, respectively. If the parent has lung cancer, the correlation is stronger (r = 0.71). On average, cancer is manifested in offspring earlier than in parents (57 and 63 years, respectively); the differences in the father-daughter and mother-son pairs are 8.2 and 2.8 years, respectively. The best prognostic parameter is the AM of cancer in the father with respect to the AM in the son (byx = 0.45).

Multiplex methylation-sensitive PCR was employed in studying the methylation of CpG islands in the RB1, p16/CDKN2A, p15/CDKN2B, p14/ARF, CDH1, HIC1, and N33 5' regions in non-small cell lung cancer (51 tumors). Methylation was observed for the two suppressor genes involved in controlling the cell cycle through the Cdk-Rb-E2F signaling pathway, RB1 (10/51, 19%) and p16 (20/51, 39%). The highest methylation frequencies were established for CDH1 (72%) and HIC1 (82%). The CpG islands of p14 and p15 proved to be nonmethylated. At least one gene was methylated in 90% (46/51) tumors and no gene, in 10% (5/51) tumors. In addition, the genes were tested for methylation in peripheral blood lymphocytes of healthy subjects. Methylation frequency significantly differed between tumors and normal cells in the case of RB1, p16, CDH1, HIC1, and N33. Gene methylation frequency was tested for association with histological type of the tumor and stage of tumor progression. Methylation index of a panel of tumor suppressor genes was established for groups of tumors varying in clinical and morphological parameters.

Polymorphic alleles of CYP17 and CYP19, which are involved in estrogen biosynthesis, were tested for association with breast cancer (BC). Microsatellite (TTTA)n and 3-bp deletion of CYP19 and single-nucleotide polymorphism T27C of CYP17 were analyzed in 123 BC patients and 119 healthy women. Of the six (TTTA)n alleles observed, allele (TTTA)8 proved to be associated with BC (11.8% vs. 6.3%, P = 0.04). Genotype A2/A2 of CYP17 was also associated with BC (32.5% vs. 20.2%, P = 0.04). Risk of BC was especially high in the presence of both factors (7.3% vs. 0%, P < 0.01). Allele (TTTA)8 and genotype A2/A2 were assumed to be risk factors of BC.

The evaluation of mutations in the p53 gene is of a big value for the laboratory diagnostics in oncology. The p53 gene was analyzed, within the present case study, in blood plasma, in which tumor-origin DNAs were detected, as well as in blood cells and in tumor tissue. Optimal conditions were defined for polymerase chain reaction (PCR) with primers. The adequacy of the conditions selected for PCR and for SSCP-electrophoresis, when used in the analysis of mutations in exons 5-8, gene p53, was shown by the method of direct sequencing. The first results on revealing the frequency of gene-p53 mutations were obtained for some of the oncology diseases in the studied population.

Expression of the markers was studied immunocytochemically on cytological preparations of breast tumor, ploidy was assessed by flow cytofluorimetry. The material was divided into 2 groups: diploid--41.4% cases and aneuploid--58.6%. Hyperexpression of Her-2/neu in the first group was observed in 54.5%, in the 2nd group in 48.6% cases. Expression of Ki-67 in 63.4% and in 68.9%, respectively. Tumours with high proliferative activity were numerous in aneuploid tissues, and in diploid tumours moderate activity prevailed (p = 0.006). Direct correlation between the markers was observed, high expression of Ki-67 more frequently was associated with positive expression of Her-2/neu. Thus, aneuploid tumours with high proliferative activity and hyperexpression of Her-2/neu are more aggressive tumours of a large size.

78 kidneys removed for cancer in 1999-2002 were studied for multifocal growth of the tumor. The results demonstrated a direct correlation between multicenter growth of renal cell carcinoma (RCC), size of the primary tumor and stage of the disease. Most of the additional foci were detected at the distance of more than 2.0 cm from the primary tumor, i.e. outside the safety zones in conduction of organ-saving operations. Frequent coincidence of a morphological type of the primary and additional tumors (70%), degree of cell differentiation (80%) and higher incidence of multifocality in infiltrative growth of the primary tumor support the existing hypophysis about the role of intra-organ metastasizing as the underlying cause of multifocal lesions. Also, the study confirmed low preoperative detection of multifocality: 1.8% preoperatively and 19.2% under investigation.

We estimated ploidometric nuclear parameters of predominating cell population in various prostatic adenocarcinomas in using WHO vs Glison classification to define advantages of the above classifications. Ploidometry provides information on cell composition and proliferative activity of cells. We observed greater ploidity in different grades of cell atypia. WHO histological classification by differentiation grade corresponds to ploidometric characterization of tumor cell populations but fails to reflect the presence of different histological types of the tumor. Currently introduced classification by Glison is more effective both diagnostically and prognostically as it reveals a dominant histological type of the tumor and shows other variants of tumor growth progression.

Colon cancer is one of the most widespread pathologies with high mortality due to recurrence and metastasis. The molecular methods of diagnosis, prognostication and biotherapy in colorectal cancer enjoyed a rapid progress during the recent decades. The hypothesis on the key role of impaired intercellular gap junctions in the onset and progression of malignant tumors is a promising trend in carcinogenesis research. We have recently discovered a variety of tumor-specific mutations of connexin 43 gene in advanced colorectal cancer (Oncogene. -2002.-Vol.21, No.32-pp.4992-4996), which confirms the above hypothesis in malignant tumor progression. We believe that further studies of connexins' mutation changes in tumor growth is a promising trend in research of its etiology and pathogenesis and in designing new methods of diagnostics and treatment of colonic and other gastrointestinal cancers.

DNA-diagnosis became to be a mandatory element of the modern oncological care. The molecular-genetic approaches aimed at identifying the high-risk categories in oncology are mostly-renowned. They comprise the diagnosis of the so-called familial cancerous syndromes, the identification of unfavorable normal genome variations (gene polymorphisms) and the detection of a number of oncoviral infections. Laboratory procedures facilitating the individualization of treatment schemes and the monitoring of oncology patients have been widely introduced into clinical practice. The turn between the 20th and 21st centuries was marked by the emergence of principally new DNA-analysis methods designed for a comprehensive and integral genome evaluation. The application of such radically novel techniques in the molecular diagnostics essentially promoted the detection and treatment of neoplasms.

Namalwa cells originating from the malignant human lymphoma have been analyzed cytogenetically upon short-time exposure to subtoxic doses of inhibitors of DNA replication and synthesis, either etoposide or fludarabine. The intact cells were characterized by the modal class of the chromosomes within the diploid range with the proportion of the aberrant cells amounting to 16.0 +/- 0.5%. Upon exposure to etoposide the percentage of the aberrant cells increased amounting to 26.1 +/- 2.9 through 39.8 +/- 1.7% depending on the duration of the exposure and the dose of the drug. At the same time the number of the polyploid cells increased but the modal class retained within the diploid range. Upon exposure to fludarabine the percentage of the cells with the aberrant chromosomes increased to 57.1 +/- 2.9%. Two modal classes appeared--the first approaching the diploid number and the second being polyploid. The exposure to either etoposide or fludarabine resulted in increasing number of the chromatide aberrations with more frequent involvement of #1, #2, #5, #6, #7, #11, #13, #14, #16 and #17 chromosomes. The data obtained have shown the susceptibility of Namalwa cells to the subtoxic concentrations of the inhibitors of DNA synthesis and replication used in the study resulting in the survival of the novel clones resistant to the drugs.

Human multiple myeloma (MM) cell lines are widely used to investigate chromosome rearrangements typical for this disease. However, during cell cultivation both numeral and structural chromosome rearrangements usually take place in addition to changes of structural and functional status of particular chromosome regions. We investigated karyotype and morpho-functional status of nucleolar organizer regions (NORs) in human cell line MM U-266. Cytogenetic analysis (G-banding) showed karyotypic stability and balanced chromosome set in hypodiploid (n = 44) U-266 cells. We found the presence of rearranged chromosomes, typical for U-266, which retained throughout many year cell cultivation. At the same time, further chromosome rearrangements were shown, along with a tendency of cells to polyploidization. Using FISH (rDNA-probe) and AgNOR-staining techniques, we found that only 4 of 8 NORS were Ag positive (AgNOR), whose dimensions varied from 1 to 3 units of arbitrary scale (u.a.s.). The average summarized AgNOR size was 7.19 +/- 0.03 u.a.s. Peculiarities of the NOR morpho-functional status in U-266 cells are discussed.

To evaluate efficacy and tolerance of glivek in chronic myeloid leukemia (CML) in patients who failed interferon-alpha (If-a) preparations.

Prostatic biopsies at different stages of carcinogenesis from 75 patients were studied. Increase of ploidy indices was established as carcinogenesis progressed. Characteristics of ploidy found in this investigation may be used for improvement of biopsy morphological studies.

This study for the first time demonstrates a physical and functional interaction between the Ca(2+)-binding protein Mts1/S100A4 and tumor suppressor p53 protein. Using different in vitro and in vivo approaches, we have found that Mts1 can bind to the C-terminal regulatory domain of p53. The Mts1 binding to p53 promotes activation of the reporter gene transcription in vivo. A modulation of the p53 target gene (p21/WAF, bax, mdm-2, and thrombospondin-1) expression was observed upon Mts1 induction in the cells expressing the wild-type p53. These results suggest that the ability of Mts1 to enhance p53-dependent apoptosis of tumor cells leads to the decrease/disappearance of the tumor cells expressing the wild-type p53. Thus, Mts1 promotes selection of more aggressive, metastatic phenotype during tumor progression.

We studied the role of nitric oxide synthase during tumor growth in oncovirus-induced tumor mutants of Drosophila melanogaster. The lines with different capacity for malignancy differed reliably in the level of enzymatic activity. It was shown using specific inhibitors of neuronal and inducible isoforms that the neuronal isoform was not involved in tumor formation, while the inducible one appears to play an important role in tumor growth inhibition. This isoform was identified with the help of immunoblotting and monoclonal antibodies against inducible nitric oxide synthase.

Multiplex methylation-sensitive PCR was employed in studying the methylation of CpG islands in the RB1, p16/CDKN2A, p15/CDKN2B, p14/ARF, CDH1, MGMT, HIC1, and N33 promoter regions in breast carcinoma (105 tumors). Methylation was often observed for the two major suppressor genes involved in cell-cycle control through the Cdk-Rb-E2F signaling pathway, RB1 (18/105, 17%) and p16 (59/105, 56%); both genes were methylated in 13 tumors. Methylation involved p15 in two (2%) tumors; CDH1, in 83 (79%) tumors; MGMT, in eight (8%) tumors, and N33, in nine (9%) tumors. The p14 promoter was not methylated in the tumors examined.

Accumulation of genetic and epigenetic aberrations leads to malignant transformation of normal cells. Functional studies of cancer using genomic and proteomic tools will help to reveal the true complexity of the processes leading to cancer development in humans. Until recently, diagnosis and prognosis of cancer was based on conventional pathologic criteria and epidemiological evidence. Certain tumors were divided only into relatively broad histological and morphological subcategories. Rapidly developing methods of differential gene expression analysis promote the search for clinically relevant genes changing their expression levels during malignant transformation. DNA microarrays offer a unique possibility to rapidly assess the global expression picture of thousands genes in any given time point and compare the detailed combinatory analysis results of global expression profiles for normal and malignant cells at various functional stages or separate experimental conditions. Acquisition of such "genetic portraits" allows searching for regularity and difference in expression patterns of certain genes, understanding their function and pathological importance, and ultimately developing the "molecular nosology" of cancer. This review describes the basis of DNA microarray technology and methodology, and focuses on their applications in molecular classification of tumors, drug sensitivity and resistance studies, and identification of biological markers of cancer.