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S Saez

Publications and source records attributed to S Saez.

At least 19 recordsLinked to original sources
PubMed1995

Influence of tumor derived fibroblasts and 1,25-dihydroxyvitamin D3 on growth of breast cancer cell lines.

Fibroblasts are known to be present in variable amounts in human breast adenocarcinoma tissue. In order to investigate if they influence in some way the proliferation rate of the carcinoma cells, we developed a coculture model in which cells of well characterized breast epithelial cell lines were seeded and grown in microchamber slides along with fibroblasts derived from breast tumor biopsies. As representatives of hormone dependent and independent tumor cells, we used MCF-7 and BT-20 cell lines. A third line, NPM-21T, derived from non proliferating mastopathy cells immortalized by SV-40 T DNA transfection, was representative of non tumor epithelial cells. The proliferation rate of the adenocarcinoma and epithelial cells was assessed by measurement of the BrdU labeling index, the cells being identified by specific beta-actin immunostaining. It was found that the proliferation of the adenocarcinoma cells was significantly increased in the presence of fibroblasts, while that of immortalized cells was not. Moreover, 1,25(OH)2D3, which was known to be a negative regulator of carcinoma cell growth, was found to be able also to blunt the overgrowth in the presence of fibroblasts. The absence of response of NPM-21T cells to the presence of fibroblasts suggests that the tumor cells could be the origin of their own overgrowth, through an indirect mechanism mediated by the fibroblasts. The factors which are involved and the 1,25(OH)2D3 mechanism of action are not yet identified.

Adenocarcinoma↗
PubMed1995

[Medical adverse effects of chemoprevention: the example of tamoxifen].

Tamoxifen, a synthetic antiestrogen widely used for the treatment of breast cancer, is also being proposed for the prevention of this cancer among women at high risk for the disease. Such an approach requires an objective and accurate evaluation, not only of the expected beneficial effects, but also of the potential iatrogenic side effects which could result from the administration of this drug to a population of healthy women. The present article summarizes our present knowledge which results from studies, all carried out on women with breast cancer: we review the side effects on the female genital apparatus, the ovarian and non ovarian endocrine side effects, the effects on cardio-vascular and thromboembolic risks, on bone metabolism and on ocular and hepatic side effects. The potential carcinogenic role is only briefly mentioned since it has already been the subject of a publication in this journal.

Adult↗
PubMed1995

[Breast cancer, model in biological and clinical oncology. Role of hormones and growth factors].

It is shown that breast cancer can be considered as a paradigm of comprehensive cancer biology and treatment based on analysis of each individual tumor. Historically, it was first observed in breast cancer that tumor progression could be dependent on factors regulating physiological activities (i.e. hormones). It was also observed that tumor growth progressively becomes autonomous, through successive degradation of the multiple steps involved in the control of cell proliferation and differentiated activities. Hormonal treatment of breast cancer provided the first example of targetted biotherapy. The discovery of the mechanism of action of hormones via specific receptors opened the field of tumor tissue analysis to determine the main biological factors involved in tumor progression. In the near future, such data should be the best guide for selection of the most appropriate treatment in each individual case.

Breast Neoplasms↗
PubMed1994

Analysis of epidermal growth factor receptor mRNA expression by polymerase chain reaction assay in 94 human breast adenocarcinoma tumors.

It is well known that breast cancer cells can synthesize and secrete various growth factors that are able to stimulate tumor growth through autocrine and/or paracrine mechanisms. EGF is one of these growth factors involved in normal breast epithelial development and tumor proliferation. EGF and TGF alpha (EGF-like peptide) are produced in variable amounts and both bind to the EGF receptor (EGF-R). Previous investigation in the laboratory measuring free and occupied EGF-R sites by differential ligand binding assays had demonstrated that non-occupied and total binding sites were present in 54 and 90% of 216 breast tumor biopsies respectively. EGF-R appeared to be totally masked by endogenous ligand in 40 and 21% of estrogen receptor positive and negative tumors respectively. The aim of the present study was to check by a molecular method the expression of the EGF-R gene. The PCR method was applied to 94 tumor samples of the previous series. Total RNA was treated with 0.5 units of Rnase-free Dnase/mg of RNA to remove any contaminating DNA. We simultaneously reverse transcribed and amplified another transcript (beta-actin) as an internal standard. Both signals were present in 88 of the 94 samples while the presence of EGF-R was detected in 74 of them when assessed by radioligand assay. The findings indicate that 93% of the tumors analysed in this series expressed EGF-R mRNA, in agreement with our previous data on occupied EGF-R sites, i.e. two-fold more than by using the standard binding assay. No significant correlation was observed between the expression of the EGF-R gene and the estrogen receptor content.

Adenocarcinoma↗
PubMed1993

William L. McGuire Memorial Symposium. 1,25(OH)2D3 modulation of mammary tumor cell growth in vitro and in vivo.

The biological role of 1,25(OH)2D3 in controlling Ca++ homeostasis in the body has been identified and widely investigated for a long time. More recently its effect in regulating cell proliferation or differentiated activity was described in a variety of normal and malignant cells. The present study was carried out to investigate the different aspects and biological mechanisms of this activity and to determine if the use of 1,25(OH)2D3 in the treatment of breast cancer patients could be considered. It is found that 1,25(OH)2D3 reduces the proliferation of MCF-7 and BT-20 cells lines regardless of their sex steroid receptor status. This effect is related to the concentration, from 10(-12) M to 10(-8) M. Its amplitude is less in other cell lines, but it opposes the EGF-induced increase of proliferation. It is observed that the proliferation rate of MCF-7 and BT-20 cells is increased when these tumor cells are cocultured with fibroblasts derived from breast tumor biopsies and that 1,25(OH)2D3 reverses this process. Moreover, experiments on DMBA induced mammary tumors in Sprague Dawley rats found that 1,25(OH)2D3 given at non toxic doses reduces significantly the tumor proliferation. These data showed that 1,25(OH)2D3 at low doses is effective on the proliferation of BT-20 and MCF-7 cells and on the paracrine growth stimulatory effect observed in the presence of fibroblasts. They suggest that 1,25(OH)2D3 or related synthetic molecules which are less active on Ca++ metabolism could be useful in the treatment of breast cancer patients.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1992

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, E.C. 1.2.1.12.) gene expression in two malignant human mammary epithelial cell lines: BT-20 and MCF-7. Regulation of gene expression by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3).

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the control of glycolysis. Its gene expression was analyzed in two breast cancer cell lines of human origin, BT-20 and MCF-7. We used a cDNA probe of 1.3 kb for Northern blot hybridization. It is found that GAPDH mRNA is overexpressed only in the poorly differentiated BT-20 cell line and that treatment of these cells by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) stimulates GAPDH mRNA expression in a dose-and time-dependent manner. The present investigation on the BT-20 cells indicates that the expression of GAPDH is sensitive to 1,25-(OH)2D3 and up-regulated by low doses of this steroid.

Breast Neoplasms↗
PubMed1992

[Cancer of the breast at the time of diagnosis. A national CANAM study: analysis of 3007 cases].

Between April 1988 and February 1990, 3,007 cases of female breast cancer were recorded among people insured by CANAM*; 118 cancers were bilateral from the start and 2,889 were unilateral. At the time of diagnosis the patients' age ranged from 24 to 101 years (median: 60 years), and 35.6 percent of the tumours were virtually subclinical. Lymph node involvement was clinically absent in 75 percent of the cases and histologically absent in 57 percent. In women under 50 the proportion of small TO-T1 tumours was greater than 40 percent, which pleaded for detection before the age of 50 years. Conversely, in economically weak populations and in women who were followed up for cancerous disease, breast cancer was diagnosed at a later stage. In older (retired) women (median age: 74), who accounted for 44 percent of the whole population, the proportion of advanced tumours was practically doubled (T4: 11 percent versus 6 percent), and the probability of metastases at the time of diagnosis had risen from 4.2 to 6.1 percent. In these patients, clinical examination once a year should contribute to an earlier diagnosis.

Adult↗
PubMed1992

Measurement of occupied and non-occupied epidermal growth factor receptor sites in 216 human breast cancer biopsies.

The epidermal growth factor (EGF) is one of several growth factors involved in normal breast epithelial development and tumor proliferation. EGF and EGF-like peptide TGF alpha bind and activate the same membrane receptor protein. This receptor (EGF-R) has been recently studied in breast tumor biopsies and its detectability reported as a prognostic indicator. However, normal and tumor tissue themselves produce EGF and related peptides in variable amount. This suggests that the standard measurement of EGF-R by binding assay should reflect only the number of non-occupied receptor sites. Based on this observation, the presence of occupied sites (EGF-R2) has been assessed in 216 human mammary tumor biopsies simultaneously with the direct measurement of non occupied EGF receptor sites (EGF-R1) and the results compared to estrogen and progesterone receptor status (ER, PGR). EGF-R1 and EGF-R2 were evaluated by 2 separate (125I) EGF binding assays performed on 2 aliquots of tumor crude membrane fraction, the first one directly, the other after dissociation of the endogenously bound ligand. The validity of the method has been assessed on membrane fractions prepared from human placenta. It is shown that the dissociation does not modify the binding dissociation constant. ER and PGR were measured by the dextran coated charcoal method. Results greater than 10 fmol/mg of membrane or cytosol protein were considered as positive. It is found that EGF-R1 and EGF-R2 are detectable in 54 and 90% of the cases, indicating that EGF-R is masked by endogenous ligand in 36% of the tumors.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding Sites↗
PubMed1992

[Adjuvant therapy of breast cancer with tamoxifen and endometrial carcinoma].

Twenty cases of endometrial carcinoma are reported, observed in women who had received long term tamoxifen treatment as adjuvant therapy for breast cancer. Furthermore, a specific study was carried out on an homogeneous group of 790 women with an operable breast cancer, 318 receiving tamoxifen as adjuvant treatment. Five endometrium carcinoma were observed in the group under tamoxifen versus none in the other group. From an analysis of these cases, authors will carry out a case control study to evaluate the relative risk of endometrial carcinoma for patients under tamoxifen. They underline the importance of gynecologic follow up procedures for all women receiving this endocrine therapy.

Adenocarcinoma↗
PubMed1991

1,25-Dihydroxyvitamin D3 increases epidermal growth factor receptor gene expression in BT-20 breast carcinoma cells.

In the breast cancer cell line BT-20 which displays a high number of Epidermal Growth Factor Receptors (EGF-R), we have previously observed that 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) at low concentration (0.1 nM) significantly reduces the proliferation rate while it upregulates the EGF binding capacity. The aim of the present investigation was to analyze EGF-R mRNA expression in 1,25-(OH)2D3 treated BT-20 cells. It is found that in cells treated for several days, the EGF-R mRNA levels are increased in relation to the dose from 0.01 to 1 nM. To investigate the time course of the response, cells received the drug only once and were harvested at different times. The data suggest that the stimulation of EGF-R mRNA expression is dose- and time-dependent. Therefore, the increased EGF binding capacity previously demonstrated can be related to the increase of EGF-R transcript levels.

Blotting, Northern↗
PubMed1991

Status of sex steroid hormone receptors in large bowel cancer.

To determine the potential role of sex steroid hormones in the development of colorectal tumors in humans, specific androgen (AR), estrogen (ER), and progesterone (PGR) receptors were investigated in normal mucosa (NM) and in tumor (T) paired biopsy specimens from 94 patients. Androgen receptors were detected in 98% and 96% of NM and T samples, ER in 91% and 83% of NM and T biopsy samples, whereas PGR were detected only in 14% and 10% of NM and T specimens, respectively. These incidences are independent of the sex and age of the patients. They are not related to tumor localization, histologic grade, or stages of Dukes' classification. Scatchard analysis of labeled ligand binding indicated the existence of one single class of high affinity binding sites; the calculated dissociation constant (Kd) was 1.7 +/- 0.6 10(-9) molar concentration (M) for 5 alpha-dihydrotestosterone (DHT) and 0.6 +/- 0.3 10(-9) M for estradiol (E2). These values were identical in NM and T tissue for both AR and ER. The binding capacity for DHT was 148 +/- 67 and 93 +/- 43 fmol/mg of cytosol protein in NM and T tissues, respectively (P less than 0.05). The ER content was lower and similar in NM and T biopsy specimens: 19 +/- 9 and 18 +/- 10 fmol/mg protein, respectively. The PGR content was 10 +/- 4 in NM versus 17.5 +/- 6 fmol/mg protein in T specimens. It is observed that the elevated AR in normal mucosa is not related to any known function for androgens in the digestive tract. The receptor pattern observed in tumors does not support the hypothesis previously raised in the case of chemically induced colonic tumors in rodents.

Adenocarcinoma↗
PubMed1991

Sex steroid and 1,25-dihydroxyvitamin D3 receptors in human colorectal adenocarcinoma and normal mucosa.

In order to determine the potential role of sex steroid and 1,25-dihydroxyvitamin D3 in the spreading of colorectal cancer, previously hypothesized from epidemiological and experimental data, specific androgen (AR, n = 94), estrogen (ER, n = 60), progesterone (PGR, n = 50), and 1,25-dihydroxyvitamin D3 receptors (VDR, (n = 111) were investigated in human colorectal adenocarcinoma (AC) and compared with the normal adjacent mucosa (NM). Scatchard analysis and competition studies of binding data did not reveal any difference between the biochemical behavior (affinity, specificity, and sedimentation coefficient) of the normal and tumoral tissue receptors. For ARs and ERs, high incidences were found (92 of 94 and 90 of 94 in NM versus 46 of 60 and 40 of 60 in AC, respectively) in both classes of tissues, while they were low for progesterone (7 of 50 and 5 of 50 in NM versus AC). While for sex steroid receptors the incidences did not vary with sex and age of the patients or the location and histopathological grade of the tumor, the VDR incidence was lower in AC (35 of 111) than in NM (99 of 111) and decreased significantly from the right colon to the rectum in adenocarcinoma. Binding capacities were similar in NM and AC for ERs and VDRs, whereas AR levels in NM were significantly higher than in AC. The expression of VDRs in some colorectal tumors suggests a possible clinical significance. No known function for sex steroid receptors is related to their presence in human colorectal tissues and their pattern in carcinoma does not support any hypothesis previously raised in the case of chemically induced colonic tumors in rodents.

Adenocarcinoma↗
PubMed1991

1,25-Dihydroxyvitamin D3 receptors and human colon adenocarcinoma.

Epidemiological evidence suggests that dietary calcium and 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) might have a protective effect against colorectal cancers. Since the presence of receptors is required for steroid action, specific 1,25-(OH)2D3 receptors (RD3) were investigated in biopsies taken at different levels of the colon. The study involved 90 biopsies from patients operated on for colorectal adenocarcinoma. They were paired biopsies from adenocarcinoma tissue and adjacent normal mucosa. In addition, 26 normal intestinal mucosa biopsies from patients without cancer were examined. RD3 receptors were assayed in tissue extract by the dextran-coated charcoal technique and also characterized by sucrose density gradient sedimentation. Scatchard analysis showed a single class of specific high affinity-low capacity sites binding for 1,25-(OH)2D3. The incidence of RD3 was 86 per cent in normal mucosa (n = 77) and lower in carcinoma (n = 34), for which the incidence decreased significantly (P less than 0.001) from right colon (58 per cent) to left colon (37 per cent) and rectum (19 per cent). These data suggest that the normal colon is a potential target organ for 1,25-(OH)2D3 which might modulate calcium transport in the colon. Loss of receptivity to 1,25-(OH)2D3 is associated with malignant transformation.

Adenocarcinoma↗
PubMed1991

Effects of an androgenic derivative on pre-established mammary tumours chemically induced in the rat.

The effects were studied of an androgenic derivative--danazol--administered at doses of 10-12 mg kg-1 day-1 during 97 days to rats with dimethylbenz[a] anthracene-induced mammary tumours. Our main observations were as follows. (a) Danazol did not influence ovarian function at the end of the assay. (b) The treatment with danazol reduced the incidence (P less than 0.05), number of tumours (P less than 0.05) and volume of malignant mammary tumours; on the other hand, the values of these parameters for benign tumours and those of doubtful expression were similar in both experimental groups. (c) Such differential action of Danazol seems to be due to the different incidence and/or content of receptors of both types of tumours. (d) The latter results lead to a hypothesis for the mechanism of action of danazol based on its behaviour at different levels.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1990

Influence of experimental conditions on osteoblast activity in human primary bone cell cultures.

Primary bone cell cultures were derived from human bone explants. Cellular activity was characterized by the alkaline phosphatase (AP) activity, osteocalcin, and type I and III collagen secretions in the supernatant. The determination of bone cell activity was performed in three different wells. No significant difference was noted between wells: the coefficient of variation was 8.0 +/- 2.9% for AP activity, 18.3 +/- 1.9% for osteocalcin secretion, and 22.5 +/- 14.3% for collagen. The AP activity and osteocalcin secretion significantly decreased with the number of passages: they were the highest after the first passage. Between each subject, the coefficient of variation was 85% for AP activity and osteocalcin secretion and 63 and 57% for type I and III collagen secretion, respectively. The AP activity did not differ with the age or sex of the donor. In contrast, osteocalcin secretion was significantly lower in females than in males. In males, osteocalcin significantly decreased with the age of the donor (r = -0.61; p less than 0.05). Cellular activity did not depend on the site of the biopsy. When bone explants from one donor were cultured in two different petri dishes, the activity of cells was similar in both dishes, except in one case. Primary cell cultures derived from human bone explants are the only model providing untransformed osteoblastlike cells of human origin. Because of the experimental conditions, some factors may have influenced the cellular activity and they must be taken into account to validate further in vitro studies.

Adolescent↗
PubMed1990

Evidence of 1,25-dihydroxyvitamin D3-receptors in human digestive mucosa and carcinoma tissue biopsies taken at different levels of the digestive tract, in 152 patients.

Epidemiological and experimental data suggest that dietary calcium and 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) are protective against colorectal cancers, while their activity on colon mucosa still remains unknown. Since the presence of receptors is required for steroid action, specific 1,25-(OH)2D3 receptors were investigated in biopsies taken at different levels of the digestive tract from the oesophagus to the rectum and in pancreas. The total study involved biopsies from 152 patients. In 82% of the cases they were paired biopsies in adenocarcinoma tissue and in adjacent normal mucosa (NM). There were 120 operated on for colorectal adenocarcinoma (HCRA). 1,25-(OH)2D3 receptor was assayed in tissue extract by the dextran-coated charcoal (DCC) technique and also characterised by sucrose density gradient centrifugation. Scatchard analyses showed a single class of specific high affinity-low capacity sites binding for 1,25-(OH)2D3 with a Kd = 1.48 +/- 0.8 x 10(-10) M (n = 119). The sedimentation coefficient of the steroid receptor complex was approximately 3.2 S. The incidence of 1,25-(OH)2D3 receptors was significantly higher in NM (82.5%) than in HCRA (34.5%). In HCRA this incidence decreased from right colon (64.7%) to left colon (27.7%) and rectum (15%). All positive HCRA in left colon and rectum (16/76) were histologically well differentiated. The receptor content in NM and HCRA was in the same range: (median) 10-314 (58) and 13-175 (64) fmol/mg protein. These data suggest that 1,25-(OH)2D3 may modulate calcium transport in colon, as in the intestine. Also, loss of receptivity to 1,25-(OH)2D3 is observed as associated with malignant transformation of the human colorectal mucosa.

Biopsy↗