Microsatellite instability and loss of heterozygosity in primary breast tumours.

Allelic imbalance or loss of heterozygosity (LOH) studies have been used extensively to identify regions on chromosomes that may contain putative tumour suppressor genes. We looked for evidence of microsatellite instability (MI) and LOH on chromosome 7q, 10q, 11p and 17q using seven polymorphic microsatellite markers. In 42 paired breast cancer-peripheral blood DNA samples we identified 24 tumours (57%) exhibiting genetic alterations. Twenty-one specimens exhibited LOH (50%), while 11 specimens exhibited MI (26%) in at least one microsatellite marker. The most frequent incidence of LOH was found for the marker THRA1 (8/33, 24%) indicating that thra I gene becomes a strong candidate tumour suppressor gene, whereas of MI it was D10S109 (3/26, 12%). These MI and LOH data were analysed using a range of clinicopathological parameters. Tumours displaying MI with no evidence of LOH and tumours exhibiting MI and LOH belonging to stage II or III were found, however none were at stage I. These data suggest that MI may be an early event in mammary tumorigenesis whereas LOH occurs at a late stage. A significant association between the absence of oestrogen receptors (p < 0.01) and the absence of both oestrogen and progesterone receptors (p < 0.001) at 17q21 were observed, indicating a possible relationship between specific genetic changes at this region and hormonal deregulation in the progression of breast cancer.

Pregnancy and offspring after the appearance of breast cancer.

The objective of this study was to investigate the influence of pregnancy in breast cancer prognosis of women under the age of 35 years. Two hundred and forty-three women with breast cancer, from three oncology departments in Athens, were investigated. Twenty-one got pregnant (7.91%) 7-100 months after breast cancer diagnosis and in a median time period of 31 months. All women had mastectomy apart from 2 who had only lumpectomy as surgical procedure. Thirteen of 21 were treated with radiotherapy and 17 of 21 had also adjuvant chemotherapy mainly with CMF for 6 cycles. Sixteen children from 14 mothers were born and the rest of the patients underwent an abortion between the 2nd and 5th month of pregnancy. All children were healthy and grew up normally up to the age of 12-142 months (end of the study) and their median age of 51 months. Only 2 patients had stage III disease at diagnosis while the remaining 19 had stage I-IIb. Three cancer recurrences were observed (14.3%) after 7-84 months. One patient had a second primary-ovarian cancer 60 months after mastectomy. Recurrence rate and survival compared with those of nonpregnant women of the same age and the stages of disease were not different. To conclude: the present study indicates that healthy offsprings can be delivered from breast cancer patients, and pregnancy does not seem to play any role in tumor recurrence.

Cyclophosphamide, mitoxantrone, fluorouracil versus conventional CMF as adjuvant treatment in node-positive breast cancer patients. A Hellenic Cooperative Oncology Group Study.

362 evaluable node-positive patients with stage II breast cancer were randomized, receiving either 6 cycles of conventional CMF or 6 cycles of the combination of cyclophosphamide (500 mg/m2), mitoxantrone (Novantrone 10 mg/m2), and fluorouracil (500 mg/m2; CNF). After a median follow-up of 51 months, 64 (36%) patients relapsed in the CMF group and 60 (33%) in the CNF group (p=0.8276). By Cox multivariate analysis, tumor size, menopausal status and number of involved nodes were retained as independently significant variables. Toxicities were remarkably similar in both groups. It appears that after a median follow-up of 51 months there is no significant difference in relapse-free survival between node-positive patients with breast cancer who received either 6 cycles of the conventional CMF or the CNF combination as adjuvant treatment.

TGF-beta 1 overexpression in breast cancer.

TGF-beta 1 belongs to a family of pluripotent growth factors (TGF beta s) and has been implicated in the development and progression of human breast cancer. There are conflicting data though, suggesting that TGF-beta has the pontency both to promote and inhibit the progression of mammary neoplasia. We examined the expression of TGF-beta 1 mRNA in 24 breast carcinomas using the technique of the reverse transcription polymerase chain reaction (RT-PCR) to obtain quantitative results. Overexpression of TGF-beta 1 gene was found in 75% of the cases. We also correlated the overexpression of the TGF-beta 1 gene with clinicopathological parameters including histological grade, tumour cellularity, oestrogen receptor status (ER), progesterone receptor status (PR) and lymph node involvement. The results led us to the conclusion that the increasing ratio of overexpression related to the stage of cancer in an analogous way (P similar to 1). No significant association was identified between the ratio of overexpression and the grade, ER, PR, or lymph node involvement (r(s) = 0.5, 0.2, 0.1, 0.1 respectively; P < 0.0001) in all categories.

Ethanol and breast cancer: an association that may be both confounded and causal.

We have examined the association of alcohol intake during various life periods, by beverage category, with breast cancer risk in Greece. A hospital-based case-control study was performed in Athens, involving 820 women with breast cancer as well as 795 orthopedic patients and 753 healthy visitor controls. Relative risk patterns were very similar with either control series, which were therefore combined to increase precision of the estimates. Drinkers of beer were at significantly elevated risk for breast cancer [odds ratio (OR), 1.34 (95% confidence interval, 1.05-1.71)]. However, drinkers of other alcoholic beverages were not at increased risk. Among beer drinkers there was no dose-response, and drinkers of both beer and other beverages had a lower OR compared to drinkers of beer only. By contrast, drinkers of 3 or more glasses of alcohol per day, mostly of spirits, were at elevated risk for breast cancer [OR for 3 - < 4 glasses per day, 3.01 (1.14-7.95); OR for 4 or more glasses per day, 3.79 (1.05-13.71)]. Reported frequency of consumption was a stronger predictor of breast cancer risk than either duration-weighted total consumption or consumption before the age of 30 years. There were no coherent patterns for interaction with menopausal status, obesity or use of menopausal estrogens. The association of very low levels of alcohol intake with breast cancer risk may be due to confounding, whereas drinking 3 or more glasses of alcoholic beverages daily appears to genuinely increase breast cancer risk, perhaps by acting as a late-stage growth enhancing factor. However, the data also are compatible with a linear relationship that has no threshold.

An investigation of hostile and alexithymic characteristics in breast cancer patients.

Two measures of personality characteristics, the Personality Deviance Scale and the Toronto Alexithymia Scale, were administered to a total sample of 448 women and a subsample of 100 women, respectively, attending two breast-screening centers in Athens. The relative risk of breast cancer between women with low denigratory attitudes towards others and women with high scores in this scale was 2.31. Statistically significant relations were not noted between the mean scores of alexithymia factors and breast cancer diagnosis. Our findings point to the conclusion that breast cancer patients can be distinguished from healthy women on the basis of their cognitive activity, characterized by rare denigratory thoughts.

Expression of ras Rb1 and p53 proteins in human breast cancer.

The ras, Rb and p53 genes have been implicated in the development of human breast cancer. Qualitative or quantitative changes in the expression of the ras p21 may lead to cell transformation, and this has been previously demonstrated in breast cancer. Both the retinoblastoma protein (Rb1) and the p53 gene product appear to function as negative regulators of cell division. We have investigated the expression of ras p21, Rb1 and p53 proteins in human breast cancer patients immunohistochemically, and correlated the results with a range of clinical and pathological parameters. Ras p21 expression was elevated in 65 per cent and p53 in 23 per cent of cases. Rb1 was expressed in 58 per cent of breast cancer tissues and in 75 per cent of normal tissue. Only four patients were found to have loss of Rb1 expression and also overexpression of both p53 and ras gene products. No correlations were found between the expression of these three genes and menopausal status, histological types or tumour grade. However, a correlation was found between Rb1 loss of expression and tumour diameter (greater than 2 cms), and no lymph node metastasis. Also, a significantly higher number of p53 staining specimens were found to be overexpressing the ras gene. These results suggest that all three oncogenes are most likely involved in the development of breast cancer but that their role is complex.

Elevated expression of AP-1 activity in human breast tumors as compared to normal adjacent tissue.

The levels of AP-1 activity in human breast lesions and in adjacent normal tissue were studied by a gel retardation assay. A thirty nucleotide long consensus oligonucleotide to the adenovirus E3 gene AP-1 sequence (E3AP-1) was end labelled and reacted with nuclear extracts from breast lesions and adjacent normal tissue. A total of 20 tissue extracts (8 pairs of tumor and normal tissue from the same patient and 4 tumors) were examined. All 12 tumor tissues showed elevated levels of AP-1 as compared to the 8 normal tissues. These results suggest that the AP-1 transcription factor may play a role in breast neoplasia.

Locally advanced breast cancer: the contribution of cytotoxic and endocrine treatment to radiotherapy. An EORTC Breast Cancer Co-operative Group Trial (10792).

Patients with locally advanced carcinoma of the breast were randomized to receive either radiotherapy alone, radiotherapy + endocrine therapy, radiotherapy + chemotherapy or radiotherapy + endocrine therapy + chemotherapy. In 363 evaluable patients, time to first progression was delayed significantly by both endocrine treatment and chemotherapy, the greatest effect being achieved by the combination of endocrine treatment and chemotherapy. This effect was almost entirely due to a major effect of systemic treatment on time to loco-regional progression, for which the result is highly significant, rather than time to distant metastasis in which only a non-significant trend was observed. For survival, a trend was seen in favour of the combination of hormone treatment and chemotherapy, but this effect did not achieve statistical significance. This trial suggests that current endocrine and cytotoxic treatments are only of marginal value in improving the prognosis in locally advanced breast cancer.

Absence of in vitro mutagenicity of the fluid from breast cysts of women with macrocystic disease.

Cystic fluid from 30 Greek women suffering from macrocystic disease was tested for mutagenicity in the Salmonella typhimurium mutagenicity assay using three bacterial strains in the presence or absence of liver homogenate. None of the samples tested showed mutagenic potential in this test supporting the absence of potential carcinogens in the cyst fluids.

Thyroid disease in patients with benign and malignant mastopathy.

The incidence of thyroid disease was examined prospectively in 97 consecutive patients with breast cancer (Group I) and was compared with that of 61 women with cystic breast disease (Group II) and that of 60 control women (Group III) with no breast problems. Thyroid enlargement was found in 47%, 49%, and 22% of those in Groups I, II, and III, respectively, and treatment with thyroid hormones was taken by 9.3% (I), 8.2% (II), and 5.0% (III) of the cases. The mean thyroid-stimulating hormone (TSH) concentration in those in Group I (5.4 +/- 2.2 microU/ml) was significantly higher than in Groups II (3.9 +/- 1.9, P less than 0.01) and III (4.0 +/- 1.8, P less than 0.001), whereas thyroid microsomal antibodies were detected in 13.4% (I), 9.1% (II), and 1.7% (III); mean triiodothyronine (T3) and thyroxin (T4) concentrations were similar in the three groups. When both TSH and T3 concentrations were taken into account, 24% and 17% of the patients, respectively, from Group I were no longer in the Group III range; the corresponding figures for Group II were 13% and 23%, respectively. These results indicate that breast cancer and thyroid disease are probably related, but not in a specific way, since benign mastopathy also seems to be associated with thyroid disturbances.

Seasonal variations in the frequency distribution of breast cancer in Greek women according to the month of their birth.

The frequency distribution of patients with breast cancer according to the month of their birth was examined in 1,165 women comprising the total number of patients recorded in our cancer registry from 1975 until the end of 1982. Statistical evaluation of this material using an exact chi 2 for simple null hypothesis demonstrated the existence of two high frequency peaks corresponding to March and April in the spring and September in the autumn. These frequencies were significantly higher (P less than 0.001) than those of the remaining months. Confirmation of this finding would imply the introduction of a new variant in breast cancer epidemiology.

The incidence of positive oestrogen and progesterone receptors in breast cancer of Greek women.

Oestrogen and progesterone receptors were determined in breast tumours of 80 Greek women. Oestrogen-positive receptors were found in 84.6% of the pre (n = 39) and 85.4% of the postmenopausal (n = 41) patients while progesterone-positive receptors were found in 71.8% of these women respectively. In combination, 69.2% of the pre-menopausal had both receptors detectable, 15.4% had oestrogen-positive and progesterone-negative and 2.6% had oestrogen negative and progesterone-positive while 12.8 had no detectable receptors. This incidence in postmenopausal women was 51.2, 34.2, 0.0, 14.6% respectively. The ratio of progesterone to oestradiol receptor concentration was 7.6 in the pre- and 1.7 in the postmenopausal group (p less than 0.05). In conclusion, the incidence of positive receptors in Greek women with breast cancer is similar to that of other Western populations.