The occurrence and the type of germline mutations in the RET gene in patients with medullary thyroid carcinoma and their unaffected kindred's from Central Poland.

We aimed to investigate the occurrence and types of pathogenic mutations in the RET gene in patients with MTC of the Central Poland population and in their relatives. DNA was extracted from the peripheral blood lymphocytes of a total of 330 persons, including 235 MTC patients and 95 of their unaffected kindred's. Exons 10, 11, 13, 14, 15 and 16 of the RET gene were amplified by PCR and sequenced. Sixty-seven people were found to carry pathogenic, germline mutations in the RET gene. In exon 10, C609F, C609R and C609Y (3 families), C618G, C618F (2 families), and C620G (4 families) mutations were identified. In exon 11, C634R (8 families) and C649L mutations (1 patient) were found. Five families carried Y791F mutation in exon 13. One patient with PTC revealed the presence of a Y791F mutation. In 3 families, exon 14 of the RET gene harbored the following mutations: V804L (1 patient), E819K (1 patient) and R844Q (1 patient). In 1 family, the S891A mutation was identified in exon 15, 3 families were found to carry mutations in exon16, R912P in 1 family and M918T in 2 families. In summary, of the 235 patients affected by MTC, 46 (19.6%) carried pathogenic RET gene mutations, 1 patient with RET mutation had kidney carcinoma, and 1 had PTC. The results show the occurrence of a variety of mutations prevalent in patients with MTC in the population of Central Poland. These results may contribute to a better diagnosis of medullary thyroid carcinoma.

Low frequency of the CHEK2*1100delC mutation among breast cancer probands from three regions of Poland.

The 1100delC germline mutation of the CHEK2 gene appears to contribute significantly to the overall breast cancer incidence in some West and North European countries, but seems to be much less frequent among breast cancer patients from other regions of Europe. In the present study we found, respectively, 3/487, 1/296 and 0/279 carriers of this mutation among breast cancer patients from the East-Central, South-East and West-Central regions of Poland. Two carriers of the 1100delC mutation were found among 120 patients with bilateral breast cancer, but only one had a previous family incidence of breast cancer. We found no carriers among 182 patients with unilateral breast cancer with family history of this tumor and among 64 patients with breast cancer and a second primary tumor at an other site. We conclude that the 1100delC mutation of the CHEK2 gene contributes little to the overall breast cancer burden in Poland, including familial cases of this malignancy. Further studies are still needed to evaluate the contribution of this mutation to the development of bilateral breast tumors.

A new look at the prognostic value of the estrogen, progesterone and epidermal growth factor receptors in breast cancer tissue.

The aim of the study was to evaluate the influence of the receptor status of primary breast cancer and of a number of selected clinical and morphological patient characteristics. The receptors were determined by biochemical radiocompetitive methods. Disease free survival (DFS) and overall survival (OS) were determined by Cox proportional hazard model. The influence of ER, PR, and EGFR on patient survival was analyzed in two ways: 1) as a separate parameter of each receptor and 2) as a common parameter consisting of 8 variables of concomitant presence or absence of the receptors. The first set of analyses had shown that EGFR as an independent parameter had no prognostic value either for DFS or OS because of a lack of statistical significance. Higher ER concentrations were positive and lower concentrations were negative prognostic factors, but only for DFS. PR was always a positive prognostic factor for DFS and OS and its prognostic value increased with concentration increase. In the second analysis it was found that patients with receptor status ER+PR+EGFR+; ER-PR+EGFR-; ER+PR+EGFR-; and ER-PR-EGFR- were having better parameters of DFS and OS (relative risks for DFS or OS were between 0.22-1.16). The patients with receptor status: ER-PR+EGFR+; ER+PR-EGFR-, ER-PR-EGFR+ and ER+PR-EGFR+ exhibited a more aggressive disease course (relative risks for DFS and OS were between 1.46-3.95). Moreover, it was found that tumor size, nodal status and patient age were independent prognostic factors for DFS and OS of patients.

The presence of hereditary BRCA1 gene mutations in women with familial breast or ovarian cancer and the frequency of occurrence of these tumours in their relatives.

UNLABELLED: In 48 women with familial breast cancer as well as in 22 women with familial ovarian cancer, the presence of pathogenic mutations in BRCA1 gene were found in 35.4% and 54.6% of patients, respectively. From the patients with mutations we created two groups: the CaM--probands with breast cancer and CaOv--probands with ovarian cancer. The probands with breast cancer were younger by a mean of five years than the probands with ovarian cancer (p = 0.048). METHODS: The PCR-SSCP procedure was used to find mutations in the BRCA1 gene. Fragments suspected of mutation were subjected to nucleotide sequencing. RESULTS: In the CaM group, which consisted of 17 women with breast cancer, the following mutations in the BRCA1 gene were detected: 5382insC, T300G, 3819del5 and IVS20+60ins12. The probands of the CaM group and their relatives developed a total of 49 breast and ovarian cancers. Among all these tumours the breast cancers of the probands made up 34.7%, the breast cancers of proband relatives made up 57.1% and the ovarian cancers of probands and their relatives made up only 8.2%. The CaOv group consisted of 12 probands with ovarian cancers in whom we detected only two kinds of mutations: 5382insC and 185delAG. The probands of the CaOv group and their relatives developed a total of 38 ovarian and breast cancers. Among all these tumours the ovarian cancers of the probands made up 31.6%, the ovarian cancers of their relatives made up 34.2% and the breast cancers of the relatives 34.2% of tumours. In the probands with breast or ovarian cancer the predominant mutation was 5382insC--in the BRCA1 gene detected in 76.5%, and 91.7%, respectively. Despite the predominant presence of the same mutation in probands from both groups the ratio of the number of breast cancers to the number of ovarian cancers in their relatives differed significantly (p = 0.0003). CONCLUSION: This data shows that the presence of the 5382insC mutation in the BRCA1 gene is not always associated with the development of ovarian cancer. It is very likely that the development of ovarian cancer requires some additional factor, which was common among the familial ovarian cancer patients, and was almost inexistent among the familial breast cancer patients. On the other hand, the development of ovarian cancer at a later age than breast cancer in probands suggests that some factors exist which slow down the development of ovarian cancer or which accelerate the development of breast cancer.

Changes in the concentrations of receptors of insulin-like growth factor-I, epithelial growth factor, oestrogens and progestagens in adenomyosis foci, endometrium and myometrium of women during menstrual cycle.

Concentrations of receptors of insulin-like growth factor-I (IGF-I-R), epithelial growth factor (EGF-R), oestrogen (ER), and progesterone (PR) in the adenomyosis foci, endometrium and myometrium of women during the menstrual cycle were studied. Cell membrane levels of IGF-I-R and EGF-R, cytosol and nuclear oestrogen (ERc and ERn, respectively) and cytosol progesterone (PRc) receptors were assayed in 37 samples of adenomyosis foci, 46 of endometrium and 50 of myometrium using the radioligand method. It was found that concentrations of ER and PR in all studied tissues were significantly higher in the proliferative phase than in the secretory phases. In the endometrium a similar difference concerned only the ERn but not the ERc. Adenomyosis foci as compared to endometrium and myometrium showed the highest ERn and PRc concentrations during the proliferative and secretory phase. Adenomyosis foci and myometrium contained significantly higher concentrations of IGF-I-R in the proliferative phase than in the secretory phase. In the endometrium a menstrual cycle-related difference in concentration of IGF-I-R was not significant. In the endometrium however the concentration of EGF-R was over twice as high in the proliferative phase than in the secretory phase. No significant difference in the concentrations of EGF-R in adenomyosis foci and in myometrium during the menstrual cycle was noted. The findings show that changes in regulation of the concentrations of IGF-I-R, EGF-R, ER and PR in the uterus of women during menstrual cycles are related to the tissular localization of these receptors.

Epidermal growth factor receptor and estrogen and progesterone receptors in breast cancers of premenopausal and postmenopausal patients.

AIMS AND BACKGROUND: The purpose of our research was to investigate the relations between the levels of epidermal growth factor receptor and estrogen and progesterone receptors in breast cancers of premenopausal and postmenopausal Polish women. METHODS: Cell membrane levels of epidermal growth factor receptor (EGF-R), and cytosol (c) estrogen (ER) and progesterone (PR) receptors were determined in 221 specimens of primary breast cancers. ERc and PRc were assayed using the dextran-coated charcoal procedure, and EGF-R were assayed by the radioligand method of specific saturation of the crude membrane fraction with 125I-EGF RESULTS: A statistically significant inverse correlation was found in all tumors between the concentrations of EGF-R and ERc, whereas for PRc significance of this correlation was not fully confirmed. ERc concentrations in tumors were positively correlated with age of patients, whereas EGF-R and PRc were not. In all, in premenopausal, and in postmenopausal EGF-R-positive tumors (> or = 10 fmol/mg of membrane protein), mean concentrations of ERc did not differ significantly. These groups of tumors seem to be homogeneous with respect to ERc concentrations. In EGF-R-negative (< 10 fmol/mg of membrane protein) postmenopausal tumors, the mean ERc concentration was significantly higher than in premenopausal tumors, hence these groups of tumors appear to be heterogeneous with regard to ERc contents: CONCLUSIONS: The present findings may have significance in differentiation of postmenopausal breast cancer.

Estrogen and progesterone receptor status in breast cancer in Kuwait female population.

The levels cytosol estrogen (ERc) and progesterone (PRc) receptors were determined in 315 primary breast cancers of female Arab patients aged 23-80 years. Most of breast cancers (78%) occurred in women aged 21-50 years, and only 22% were in women aged 51-80 years. Breast cancers containing ERc and PRc concentrations in the range 5-50 fmol/mg of cytosol protein (mg c.p.) were found with similar frequency in women aged under or over 50 years (53% for ERc and 43% for PRc, respectively). On the other hand, breast cancers with ERc values of > 50 and > 100 fmol/mg c.p. were twice as frequent in women aged over 50 years as in women aged under 50 years. The frequency of breast cancers with PRc level of over 50 fmol/mg c.p. in women aged over 50 years was only half that in those aged under 50 years. In breast cancers of Kuwait Arab women the higher values of ERc (> 100 fmol/mg c.p.) and PRc (> 50 fmol/mg c.p.) were less frequent than in other populations reported in literature. The low frequency of breast cancer in postmenopausal Kuwait women is associated with low proportions of tumors with higher ERc and PRc contents. In contrast to this, data from literature indicate that in the North Western European and American populations the postmenopausal incidence rise of breast cancers is associated with increased proportions of tumors with higher ERc and PRc levels.

Relations between epidermal growth factor receptor and oestrogen and progesterone receptors in breast cancers of premenopausal and postmenopausal patients in Kuwait.

The levels of cell membrane epidermal growth factor receptor EGFR and cytosol (c) and nuclear (n), oestrogen (E) and progesterone (P) receptors (R) were determined in 132 specimens of primary breast cancers. In the tumours of postmenopausal women an inverse significant correlation was demonstrated between the concentrations of EGFR vs. ERc, ERn, and PRc while no such correlation was noted in the tumours of premenopausal women. Premenopausal and postmenopausal EGFR positive tumours (> or = 10 fmol/mg membrane protein) could be regarded as homogenous with respect to the concentration of ER and PR whose mean values were low and without being significantly different. EGFR negative tumours were heterogeneous with respect to the ER and PR concentrations. Postmenopausal EGFR negative (< 10 fmol/mg membrane protein) tumours had evidently higher mean values of ER and PR than premenopausal EGFR negative tumours, but these differences were statistically significant for oestrogen receptors only. The levels of ER and PR of premenopausal EGFR negative tumours were approximated to the corresponding levels of EGFR positive tumours.

Quality control program of steroid receptor assays: an international study.

Lyophilized calf uterine cytosol standards were prepared for quality control of estrogen receptor (ER) determination, and lyophilized cytosols and tissue powders were used for quality control of progesterone receptor (PR) analysis. Two series of four samples were analyzed either for ER or PR contents, twice within one month, by 7 laboratories in 5 countries. Coefficient of variation (CV) of the between-laboratory averages assayed in a single run of ER-positive (ER+) and PR-positive (PR+) standards varied from 29.6 to 61.8% and from 32.4 to 76.2%, respectively. All laboratories, with the exception of a single value, could recognize samples of low, medium, an high ER level, as well as a negative sample. Most laboratories evaluated properly also the level of PR samples. The average between-laboratory CV values of protein determination in the relevant standards were 23%.

[Immunocytochemical evaluation of estrogen receptors in histological specimens of primary breast cancer]. / Immunocytochemiczna ocena receptora estrogenów w preparatach histologicznych pierwotnych raków sutka.

Immunocytochemical methods (ER-ICA) by using monoclonal antibodies were applied to determine the presence of estrogen receptors (ER) in 44 primary breast cancers in women. Of this 48% of the tumours were classified as receptor positive. In these tumours the nuclei had a clearly positive heterogenous colouration. In three cases a positive reaction was also found in benign epithelial cells of the breast. ER determinations by ER-ICA method were compared with quantitative analysis carried out by using radioligand and immunoenzymatic methods. There was a strong correlation between immunocytochemical ER evaluations and quantitative methods. We also found a correlation of the menopause state and patients age and ER content.

Estrogen and progestin receptor levels in uterine leiomyomata: relation to the tumour histology and the phase of menstrual cycle.

Estrogen receptor (ER) and progestin receptor (PR) levels in the myometria and uterine leiomyomata of 26 normally menstruating women were studied. No significant menstrual cycle-related change in ER level was found in the leiomyomata or normal myometria. The ER levels in normal myometria and in cellular leiomyomata, but not in usual leiomyomata, tended to be higher in the follicular phase than in the luteal one. The "cytosolic" PR levels in cellular leiomyomata and in usual leiomyomata with no or with slight hyalinization as well as in their parental myometria were significantly lower in the luteal than in the follicular phase. This was not the case in usual leiomyomata with more intense hyalinization. The findings show that the reactivity of uterine leiomyomata to estrogens and/or progestins may be related to the histological features of the tumors. This should be realized when studying the steroid receptor levels in the tumours and possibly also when planning an endocrine therapy for the leiomyomata.

Immunoreactive and estrogen-binding estrogen receptors, and progestin receptor levels in uterine leiomyomata and their parental myometrium.

Estrogen receptor (ER) and progestin receptor (PR) levels in the myometria and uterine leiomyomata of forty-four women were studied. A radio-ligand method and an immuno-enzymatic method were used for ER measurement, and only a radio-ligand method was used for PR measurement. The leiomyomata contained significantly more PR and estrogen-binding ER than their parental myometria but not the immunoreactive ER per mg of DNA. Nuclear extracts from the myometria contained a high amount of the estrogen-nonbinding immunoreactive ER; in the leiomyomata, the bulk of this particular ER fraction was extracted with cytosol. Dissimilar distribution patterns of immunoreactive, estrogen-nonbinding ER in leiomyomata and normal myometria suggest that an impaired metabolism of ER may contribute to myoma growth.