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DNA flow cytometry as a new prognostic factor in ovarian malignancies

Nowadays, it is clear that chromosomal rearrangement represents an early step in the initiation of carcinogenesis. It alters the expression of certain genes, resulting in a change in the cell's responsiveness to normal growth control mechanisms. Karyotypic abnormalities have been observed in human cancers, and here the chromosomes that are most commonly affected contain some of the genes regulating metabolism and nucleic acid synthesis. The gain or loss of such chromosomes alters the amount of specific cell products, and if these are critical for the control of cell division and differentiation the resulting cells may achieve a proliferative advantage. Furthermore, genetic rearrangements may increase the rate of spontaneous mutation in a cell line and allow rapid progression to a high malignant phenotype. The normal human somatic cell with 46 chromosomes (23 pairs) is referred to as diploid (normoploid = 2N). Cells containing a multiplum of 2 chromosome sets are termed polyploid (x 2N). The cells with double chromosome set (complete replication) are termed tetraploid (4N). A cell with fewer or more than 46 chromosomes is described as aneuploid (hypoploid or hyperdiploid, respectively) and a cell with a balanced gain or loss of chromosomes, or with structural rearrangements, which still has 46 chromosomes, is pseudodiploid. The most frequent chromosomal number in a tumor is called modal number and it represents the cell line that is responsible for tumoral growth. DNA cellular synthesis occurs at a definite time during the cell cycle consisting of 4 phases: G IIGO, S, G2 and M phases. G 1 (2N) is the time proceeding synthesis of DNA. S, is the time when doubling of the cellular DNA content in preparation for mitosis occurs. G2 (4N) is a short period following doubling of the cellular DNA content and it precedes mitosis. M (Mitosis) is the phase where chromosomal division occurs and every mother cell will give rise to two daughter cells each with diploid number of chromosomes. Most of cells remain in rest phase (GO) and during GO and G 1 phases, each cell contains the same diploid number of chromosomes (2N). The use of flow cytometry to analyze the cellular DNA content (FCM-DNA) of human malignancies has become increasingly common. Using the automated techniques, more than 10,000 tumor cells from a patient can be studied within a few minutes. In flow cytometry, a variety of dyes that bind stoichiometrically to DNA are incorporated into tumor nuclei and their florescence upon laser or mercury excitation is measured and recorded. Currently available flow cytometer can resolve two clonal cell populations, provided there is at least a 5% difference in DNA content between them. Thus, karyotypic abnormalities of two or less chromosomes are usually not detected by this method. By flow cytometry a histogram represently the relation nuclear DNA content is obtained. The main top represents cells in G IIGO phase (2N, modal DNA value), and the small peak with the double DNA values (within the double channel numbers), represents G2 + M cells (4N). The extent of abnormal DNA or aneuploidy is usually expressed as the DNA index (DI). DI is the ratio of the DNA content of the GO/Gl cells in the abnormal population to the GO/G1 cells of a normal, diploid population. The diploid subpopulation in a