Effects of sex steroids on survival and receptor expression in ovarian epithelial tumour cells.

The factors that govern the genesis and progression of ovarian cancer remain unclear. It is thought that ovarian tumours are endocrine related and hormone dependent. We therefore investigated the effects of the sex steroids progesterone, testosterone and 17 beta-estradiol on tumour cell survival and the expression of estrogen receptors (ER) and progesterone receptors (PR) in tumour cells. The study was performed on primary cell cultures derived from patients suffering from epithelial ovarian cancer. The majority of the cells isolated expressed ER and PR to some degree, the combination ER+/PR+ was the most common. Both ER and PR expression decreased after 72-h culture, revealing an unexpectedly dynamic system. The survival rates of cells cultured in progesterone seemed to be inversely related to their PR expression. Lowering levels of 17 beta-estradiol and testosterone in cell cultures reduced cell survival, but it appears that this observation depends on factors other than ER.

Low sex steroid environment affects survival and steroid secretion of ovarian tumour cells in primary cultures.

Ovarian epithelial tumours are considered to be endocrine related. The effects of an environment with low levels of the steroid hormones 17 beta-estradiol, testosterone or progesterone on cell survival and steroid secretion were studied in primary cell cultures derived from 25 patients suffering from epithelial ovarian tumours. Tumour cells cultured in 17 beta-estradiol and testosterone showed a reduced cell survival (-10.3 +/- 2.3% and -15.6 +/- 2.7% minimum survival respectively). This reduction was inversely proportional to hormone concentrations within the range studied. No similar effect was observed in the progesterone cultures. It was found that 17 beta-estradiol was secreted from the primary cell cultures and, interestingly, the amount of 17 beta-estradiol secreted increased with increasing levels of 17 beta-estradiol in the environment. Neither progesterone nor testosterone production was observed in any of the cultures studied. It is believed that 17 beta-estradiol has an antiapoptotic effect on ovarian surface epithelial (OSE) cells. Reduction of 17 beta-estradiol in the environment may inhibit this effect, resulting in reduced cell survival. The ability of ovarian epithelial tumour cells to secrete 17 beta-estradiol suggests that epithelial ovarian tumours play an active role in altering their own hormonal environment, promoting tumour progression.

Isolation and culture of ovarian tumour cells, cytological and cell survival evaluation.

The purpose of this study was to evaluate the reproducibility and reliability of the fluorometric microculture cytotoxicity assay (FMCA). Emphasis was placed on obtaining pure tumour cell cultures which were subjected to careful cytological evaluation. Preparations of 39 ovarian tumours, malignant, borderline and benign were made, of which 37 were successfully cultured. In 34 of the 37 tumour cell cultures, the epithelial cell fraction was > 90%, and in 30 of 39 cultures the epithelial cell fraction was > 95%. Transportation within 24 h and the 72 h incubation did not change the yield or epithelial cell fraction. There was a linear relationship between fluorescence and the number of viable cells. The fluorescence increased with time, making only comparisons within each assay plate possible. The sensitivity of the method makes it possible to perform many analyses on a small amount of material. The method also makes it possible to study cells derived from all stages of the disease, including benign tumours.

Reproducibility in DNA flow cytometric analysis of breast cancer: comparison of 12 laboratories' results for 67 sample homogenates.

Flow cytometric (FCM) DNA analysis yields information on ploidy status and the S-phase fraction (SPF), variables of prognostic importance in breast cancer. The clinical value of the SPF is currently being evaluated in prospective randomized trials. The widespread use of FCM DNA analysis emphasizes the importance of reproducibility (both intra- and interlaboratory). In this study, 67 nuclear suspensions of breast cancer samples were analyzed by 12 laboratories routinely performing FCM DNA analysis in breast cancer. No general guidelines were imposed; each laboratory used its own standard protocols. For DNA ploidy status (diploid vs. non-diploid), agreement was complete for 79% (53/67) of the samples, compared with 64% (43/67) of samples when tetraploidy was considered [i.e., euploid (diploid+tetraploid) vs. aneuploid (the remaining non-diploid)]. For the SPF, pairwise comparison of the results of all 12 laboratories yielded a mean Spearman's rank correlation of 0.78 (range: 0.54-0.93). For those 39 samples being categorized in low or high SPF by all laboratories, all agreed in 14 samples (36%). Similar patterns were obtained with kappa measures, agreement being good for ploidy status (diploid vs. non-diploid; overall kappa = 0.87 and 0.74 for euploid vs. aneuploid), but moderate for the SPF [overall kappa = 0.47 (for low SPF vs. high SPF vs. "no SPF reported")]. Discrepancies were chiefly attributable to differences in the categorization of the S-phase values, rather than in FCM procedures, other critical differences being in the detection and interpretation of near-diploid and small non-diploid cell populations, the definition of tetraploidy, and the choice and execution of the method used for S-phase estimation. Based on the observations of this study, detailed guidelines for FCM analysis and interpretation of data are proposed in the Appendix. Some issues remain, however, e.g., to standardize a method for S-phase calculation and tetraploid definition.

High prevalence of cervical dysplasia in female consorts of men with genital warts.

Altogether 51 regular female consorts of men attending a venereal disease clinic for genital warts were examined using colposcopy, vaginal cytology and--when needed--surgical biopsy. Abnormal cytological smears were found in 18 out of 49 consorts (37%), which should be compared with 8 out of 124 (6%) matched female controls from a family planning clinic (p less than 0.001). Possibly premalignant lesions, i.e. atypical condylomata and/or frank dysplasia, were found in 14 (27%) out of 51 consorts. The prevalence of abnormal smears or biopsy-proven dysplasia was approximately the same in consorts with and without external warts. These findings call for close attention to the risk of development of cervical dysplasia in female consorts of men with genital warts.

Morphologic changes in ileal reservoir mucosa after long-term exposure to urine. A study in patients with continent urostomy (Kock pouch).

In patients with supravesical urinary diversion, continent ileal reservoirs utilized for urinary collection were examined by endoscopy at intervals of 1 month to 9 years after surgical construction, and biopsy specimens were obtained for light microscopy and morphometry. The gross appearance of the mucosa showed alterations in the shape of the shorter and broader finger-like villi during the first postoperative month to the subsequent very low ridges and convolutions or, in some instances, flat mucosa devoid of individual villi. Starting between 1 and 3 years after the surgical construction, endoscopically smooth areas were encountered in caudal areas of the reservoir, and the areas were mixed with islands of villous mucosa. Microscopically and morphometrically, the specimens from villous areas confirmed reduction in villous height and increase in crypt depth, whereas no changes were seen in the epithelial mitotic frequency. The number of mucus-storing goblet cells increased already within 1 month after construction. Specimens obtained from smooth areas showed alterations in the intestinal structure, with reduction of crypts, decreased height of epithelial cells, and occasional epithelial denudation. No signs of fibrosis, foreign-body reaction, or dysplasia were encountered. The constant exposure to urine leads to adaptive changes of the reservoir mucosa, resulting in a true atrophy of villi, crypts, and individual epithelial cells.

Ultrastructural and histochemical changes in ileal reservoir mucosa after long-term exposure to urine. A study in patients with continent urostomy (Kock pouch).

Mucosal biopsy specimens were obtained from ileal reservoirs used for urinary collection in patients with continent urostomy. Shortly after construction of the reservoir there was a reduction in villous height and an increase in crypt depth. After 2 to 3 years of observation, avillous areas were noted in the reservoir mucosa, mixed with islets of villous mucosa. Specimens from the avillous areas showed a marked decrease in height of superficially located enterocytes, with loss of microvilli and a reduction in cell organelles. The histochemically demonstrable activity of oxidative enzymes was significantly reduced in the epithelial cells. There was an increase in the number of goblet cells and autonomic nerve fibres in the crypt layer. The constant exposure to urine led to significant alterations of the ileal mucosa resulting in avillous areas mixed with villous remnants in which many of the intestinal characteristics both structurally and functionally disappear.

Dissemination pattern of two immunologically induced transplanted malignant lymphomas in C57 B1/6J mice.

The need for syngenic tumors to elucidate factors in lymphoma dissemination prompted us to induce new lymphomas in a series of inbred C57B1/6J mice. The method of induction, immunosuppression and simultaneous immunostimulation, had a high mortality rate, but gave us 2 transplantable lymphomas. These tumors are described and compared in the present paper, with emphasis on their dissemination patterns. Both tumors disseminated rapidly from tail transplants to lymphoid organs, lungs and liver. They were characteristically dissimilar, however, in their s.c., i.p. and i.v. transplantabilities.