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PubMed1979

The leukocyte migration inhibition response to certain breast cancer-related antigens (MCF-7 and MuMTV): their potential as discriminants.

Certain oncogenic viruses have been implicated in human breast cancer, including the murine mammary tumor virus (MuMTV) and the Mason-Pfizer monkey virus (MPMV). We have used the leukocyte migration inhibition (LMI) response to assay the response to several potential breast cancer-related antigens, including MuMTV, MPMV, and a breast cancer cultured cell line, MCF-7, in 96 breast cancer patients, in 32 women with benign breast disease, and in 67 normal women. The lowest tenth percentile of control (LMI) responses was used as the cutoff point to designate responders. Breast cancer patients showed significant responses to MuMTV (49% and to MCF-7 (50%), but not to MPMV (29%). In a paired-antigen study using MuMTV and MCF-7, 75% of the breast cancer patients responded, versus 18% of the normal women (P less than 0.0050). The potential for this assay to distinguish "normal" from "breast cancer" was analyzed using a migration index derived from discriminant analysis. The ability of the assay to discriminate "normal" from "cancer" was significant (P less than 0.001) and showed a sensitivity of detecting "cancer" of 75%. The overall responses to MuMTV and MCF-7 were analyzed with reference to certain prognostic factors, but showed no relation to age, menstrual status, estrogen receptor status, or stage of disease. The above reactions suggest that a large proportion of breast cancer patients exhibit presensitization to antigenfs found in MuMTV and MCF-7, which may be cross-reactive with antigens in the primary cancer. These responses appear to be independent of major prognostic variables. Further refinement of this assay may yield one which is more highly discriminating for breast cancer.

Adult↗
PubMed1977

Glycoprotein synthesis as a function of epithelial cell arrangement: biosynthesis and release of glycoproteins by human breast and prostate cells in organ culture.

We demonstrate that a technique is available to investigate glycoprotein synthesis in organ cultures of human breast and prostate surgical specimens where the 3-dimensional epithelial cell arrangement remains intact. Malignant breast and prostate epithelium maintained their capacity to synthesize glycoproteins for at least 3 days as followed by the incorporation of [3H]glucosamine into macromolecules. Over 70% of incorporation was by malignant cells as judged by autoradiography. Labeled glycoproteins were released into glandular lumina and consequently into the culture fluid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed predominantly one group of macrmolecules released with an apparent molecular weight of 48,000 +/- 6,000 daltons. This glycoprotein was found in all of the breast specimens studied, which included 1 medullary, 1 infiltrating lobular, and 8 infiltrating duct carcinomas. The pattern was independent of the availability of estrogen receptors. A similar glycoprotein was also observed in the culture media from a Grade I and a Grade II well-differentiated infiltrating prostate carcinoma. Incorporation was below the level of detection in 4 of 6 cases of benign prostatic hyperplasia. A more complex pattern of labeled glycoproteins was found in the media of a Grade II and a Grade III poorly-differentiated prostate carcinoma. The established human mammary carcinoma cell line MCF-7 synthesized and released a similar 48,000 molecular weight glycoprotein but additional components with larger molecular weights were also released. An intriguing interpretation that 3-dimensional tissue integrity restricts some glycoprotein synthesis is discussed. Cells grown in 2-dimensional monolayers could escape from such a topographic restriction and express additional families of glycoproteins.

Adenocarcinoma↗
PubMed1978

Immunologic relationship between breast carcinoma and benign breast disease as detected by the leukocyte migration inhibition assay.

Patients with benign diseases of the breast reacted in a migration inhibition assay with extracts of breast cancer and benign breast lesions and a human breast cancer-derived cell line, MCF-7. The incidence of reactivity of the patients with benign breast diseases against these antigens was similar to that of breast cancer patients. In addition, patients with breast cancer reacted to some extracts of benign breast lesions. The reactivity occurred in patients with several different histopathologic types of breast lesions, but was not found in women with no detectable pathologic lesions.

Adenofibroma↗
PubMed1978

Phytoestrogen interaction with estrogen receptors in human breast cancer cells.

The interactions of phytoestrogens with estrogen receptors were studied in the human breast cancer cell line, MCF-7. The compounds tested were coumestrol, genistein, and formononetin and the mycotoxins, zearalenone and its reduced derivative, zearalenol. All but formononetin compete for binding of [3H]-estradiol to unfilled cytoplasmic estrogen receptor or unfilled nuclear estrogen receptor sites. Relative binding affinities are zearalenol HMP (high melting point isomer) greater than zearalenol LMP (low melting point isomer) greater than zearalenone = coumestrol greater than genistein greater than formononetin. Dissociation constants estimated from competition curves show that binding affinities are high. In contrast to estradiol, phytoestrogens bind only weakly to sex steroid-binding globulin; they also do not bind to corticosteroid-binding globulin. These compounds translocate the cytoplasmic estrogen receptor and bind to unfilled nuclear estrogen receptors in whole cells. Bound nuclear receptors are then processed in a manner similar to estradiol in a step which rapidly decreases total cellular estrogen receptors. The phytoestrogens are also biologically active; they can markedly enhance tumor cell proliferation. In sum, phytoestrogens interact with the estrogen receptors of human breast cancer cells in culture and, therefore, may affect estrogen-mediated events in these cells.

Breast Neoplasms↗
PubMed2026

Screening of Estrogenic and Antiestrogenic Effects of Estradiol, Bisphenol A, and Fulvestrant Using 2D and 3D Breast Cancer Cell Systems With a Luciferase Reporter Gene Assay.

Endocrine-disrupting chemicals (EDCs) like bisphenol A (BPA) pose health risks by interfering with hormones. This study develops and utilizes in vitro 2D and 3D cell models to evaluate the estrogenic and antiestrogenic properties of compounds. Human breast cancer cell lines T47D and MCF7, stably transfected with a luciferase reporter gene (ERE-LUC), were first compared in 2D. Due to the significantly higher sensitivity and responsiveness observed in the T47D line during preliminary 2D screenings, this cell line was exclusively selected for the development of the 3D spheroid model. Cells were treated with 17β-estradiol (E2), BPA, and Fulvestrant (FUL) to assess cell viability and luciferase activity. In 2D models, T47D ERE-LUC cells showed higher responsiveness than MCF7 ERE-LUC, which failed to show significant luciferase induction with E2. In the 3D T47D model, cells exhibited significant and robust changes in luciferase activity in response to E2 and BPA, highlighting the enhanced fidelity of 3D cultures in replicating tissue conditions compared to their 2D counterparts. The study highlights the effectiveness of 3D models over 2D in evaluating estrogenic activity. Specifically, the 3D T47D ERE-LUC system serves as a superior, sensitive, and reliable platform for screening EDCs, offering benefits in cost, data speed, and reduced in vivo reliance.

Humans↗
PubMed1978

Actinomycin D prevents nuclear processing of estrogen receptor.

This communication describes a novel effect of actinomycin D (AcD) in inhibiting the nuclear processing or turnover of estrogen receptors in the human breast cancer cell line. MCF-7. In the absence of AcD, estradiol treatment results in rapid (5 min) hormone binding and translocation of unfilled cytoplasmic receptors (Rc) and binding of unfilled nuclear receptors (Rn). Thereafter, filled nuclear receptors (RnE) progressively deplete and, by 3 to 5 h, 70% are lost or processed. We now show that 1 to 2 micrometer AcD or chromomycin A3, both of which intercalate at G-C base-pairs on DNA, selectively and completely block RnE processing. In contrast, estrogen binding, translocation of receptor complex, and RnE accumulation in the nucleus are completely insensitive to AcD inhibition. At 1 to 2 micrometer, all other intercalators and inhibitors tested, including other inhibitors of transcription and replication, or inhibitors of translocation or of other functions, fail to prevent binding, translocation, or the nuclear processing step. AcD intranslocation, or the nuclear processing step. AcD inhibition of RnE processing is dependent on dose; at lower doses (100 nM decreasing to 1 nm), progressively greater RnE depletion occurs. AcD completely prevents RnE depletion if given together with or within 30 min after estradiol; at any time between 30 min and 5 h after estradiol, the processing of RnE is stopped instantly by addition of AcD. Because of the complexity of actinomycin action, several mechanisms can be proposed to explain its effect on nuclear ER levels.

Breast Neoplasms↗
PubMed1979

Glycosaminoglycans of normal and malignant cultured human mammary cells.

Glycosaminoglycans have been characterized from a normal human breast cell line (HBL-100) and two different cell lines from human breast carcinoma (MDA-MB-231 and MCF-7). The glycosaminoglycans were labeled by exposure of cell cultures to [3H]glucosamine and [35S]sulfate and then isolated from both spent media and cells by pronase digestion and cetylpyridinium chloride fractionation. They were further characterized by (a) hexosamine composition, (b) controlled-pore glass exclusion chromatography, (c) reactivity with specific enzymes (hyaluronidase chondroitinase, heparitinase, and heparinase), (d) nitrous acid degradation, and (e) DEAD-Sephadex chromatography. The results indicate that the HBL-100 line synthesizes mainly hyaluronic acid, most of which is secreted into the medium. Chondroitin sulfate and heparan sulfate are the predominant glycosaminoglycans synthesized by the cancer lines; both are found mainly in the spent medium, but the hyaluronic acid synthesized by the MDA-MB-231 line remains cell associated. The cell-associated heparan sulfate had a molecular weight in excess of 13,000 and may contain linkages susceptible to testicular hyaluronidase. The MCF-7 cells produce significantly lower amounts of glycosaminoglycans than do the other two lines.

Breast↗
PubMed2026

Microalgae-Mediated Synthesis of Gold Nanoparticles from Indonesian Chlorella vulgaris InaCC M205 with Potential Anticancer Properties for Biomedical Application.

Sustainable nanomaterial synthesis has emerged as a critical strategy to reduce the environmental burden associated with conventional chemical synthesis method. Microalgae-derived biomolecules offer a promising platform for the green production of metal nanoparticles due to their rich bioactive compounds capable of acting as natural reducing and stabilizing agents. Here, we report the eco-friendly synthesis of gold nanoparticles (AuNPs) using extract of Indonesian microalga Chlorella vulgaris extract. To optimize the synthesis process, the effects of precursor-to-extract ratio, temperature, and incubation time were evaluated. Optimal synthesis of C5-AuNPs was obtained at 37 °C for 20 h with precursor to extract ratio of 6:4, resulting in moderately stable C5-AuNPs characterized by a surface plasmon resonance (SPR) peak at 541 nm. Furthermore, Fourier-transmission infra-red (FT-IR) analysis revealed the involvement of functional groups of C. vulgaris extract in the interaction with Au+ during the production of C5-AuNPs. Transmission electron microscopy (TEM) demonstrated the formation of uniformly spherical nanoparticles with an average diameter of approximately 8.8 nm. Biological evaluation showed that the synthesized C5-AuNPs exerted pronounced dose-dependent cytotoxicity against MCF-7 breast cancer cells with an IC50 threshold of 21.17 ppm, while no toxicity appears in normal HEK293 cells. Mechanistically, the C5-AuNPs induced early apoptosis and inhibit cell-cycle progression at the stage of G0/G1. Collectively, these findings demonstrate that C. vulgaris-mediated AuNPs represent a promising preliminary in vitro findings for cancer therapy candidate.

Gold↗
PubMed1977

Effects of estrone, estradiol, and estriol on hormone-responsive human breast cancer in long-term tissue culture.

The effects of estrone, estradiol, and estriol on MCF-7 human breast cancer are compared. In this estrogen-responsive cell line, all three estrogens are capable of inducing equivalent stimulation of amino acid and nucleoside incorporation. Estriol is capable of partially overcoming antiestrogen inhibition with Tamoxifen (lCl 46474), even when antiestrogen is present in 1000-fold excess. Antiestrogen effects are completely overcome by 100-fold less estriol. Studies of metabolism of estrogens by MCF-7 cells revealed no conversion of estriol to either estrone or estradiol. All three steroids bind to a high-affinity estrogen receptor found in these cells. The apparent dissociation constant is lower for estradiol than for estrone and estriol, but all three bind to an equal number of sites when saturating concentrations are used. Tritiated estrogens used in binding studies were shown to be radiochemically pure. We conclude that estriol can bind to estrogen receptor and stimulate human breast cancer in tissue culture. Our data do not support an antiestrogenic role for estriol in human breast cancer.

Binding, Competitive↗
PubMed2026

Whole-genome analysis of Brevibacterium sanguinis AZMABM HM27: a bacterial isolate from the sea anemone Radianthus magnifica and exhibiting promising multi-therapeutic properties.

BACKGROUND: The marine anemone Radianthus magnifica harbors symbiotic microbes with promising biomedical potential, yet their diversity and therapeutic properties remain underexplored. This study aimed to characterize a symbiotic bacterium isolated from R. magnifica collected from Samalona Island, Indonesia, and to evaluate its multi-therapeutic potential. METHODS: Strain AZMABM HM27 was characterized using whole-genome sequencing, functional annotation, biosynthetic gene cluster prediction, molecular docking, and in vitro bioactivity assays. RESULTS: Phylogenetic and genome-based analyses confirmed AZMABM HM27 as Brevibacterium sanguinis, with an OrthoANI value of 97.37% and a dDDH value of 76.50% against the type strain. The genome comprises a 3,834,082 bp chromosome encoding 3,362 protein-coding genes, including 95 genes involved in secondary metabolite biosynthesis. Five biosynthetic gene clusters were predicted, including those associated with ectoine, terpene, and siderophore production. The crude extract demonstrated antioxidant activity (IC₅₀ = 0.87 mg/mL), anti-inflammatory activity (up to 60% inhibition), antidiabetic activity through α-glucosidase inhibition (up to 40% inhibition), and dose-dependent antiproliferative activity against MCF-7 breast cancer cells (74.10% viability at 1 mg/mL). Molecular docking identified a lead compound, 8,9,9,10,10,11-hexafluoro-4,4-dimethyl-3,5-dioxatetracyclo [5.4.1.0(2,6)0.0(8,11)] dodecane, with strong binding affinities to selected therapeutic targets. CONCLUSIONS: B. sanguinis AZMABM HM27 represents a marine symbiotic strain associated with R. magnifica and a promising source of bioactive compounds with antioxidant, anti-inflammatory, antidiabetic, and antiproliferative potential. Further purification, structural elucidation, and in vivo studies are warranted to validate its therapeutic potential.

Animals↗
PubMed1977

Relationship of human natural lymphocyte-mediated cytotoxicity to cytotoxicity of breast-cancer-derived target cells.

Mononuclear cells from 115 individuals were tested in a 4-h chromium release assay against two breast-cancer-derived cell lines, G11 and MCF-7, and a myeloid line, K-562, shown previously to be sensitive to natural cytotoxicity. These data were analyzed in a manner designed to detect hyperreactivity against the breast cell lines relative to the level of reactivity against K-562. A high proportion of breast cancer patients were found to be relatively hyperreactive against G11 (12/18 or 67%) and against MCF-7 (10/18 or 56%). Fibroadenoma patients were very similar to the normal females, with 0/11 hyperreactive to G11 and 1/11 (9%) to MCF-7. However, several normal males (7/17 or 41%) were hyperreactive to G11 but not to MCF-7 (2/17 or 12%). Colon cancer and lung cancer patients were also more hyperreactive to G11, 4/8 or 50% and 4/6 or 67%, respectively, than they were to MCF-7, 1/8 or 13% and 1/6 or 17%, respectively. Only fibrocystic patients resembled the breast cancer patients, with some but not as many individuals being hyperreactive to G11 (3/8 or 38%) and to MCF-7 (2/8 or 25%). With another group of individuals reproducibility of the method was demonstrated, with only 1/14 or 7% of normal females and 12/17 or 70% of breast cancer patients being hyperreactive to G11. Thus, natural cytotoxicity toward K-562 can be related to breast cancer-associated cytotoxicity toward MCF-7 in a way that distinguishes a majority of breast cancer patients specifically from other groups of individuals.

Breast Neoplasms↗
PubMed1979

Content of gonadotropins in cultured human malignant cells and effects of sodium butyrate treatment on gonadotropin secretion by HeLa cells.

Twenty-one of 82 human cell lines examined for production of human chorionic gonadotropin and its subunits (HCG-alpha and HCG-beta) produced either one or both subunits at some phase in their growth. Of these, 14 produced an excess amount of free alpha subunit, and seven produced HCG-beta or complete HCG without evidence for free alpha subunit synthesis. Five of the HCG-producing cell lines also contained or secreted the beta subunit of human luteinizing hormone. CBT cells derived from a glioblastoma multiforme and JAR choriocarcinoma cells secreted significant amounts of the beta subunit of human luteinizing hormone, while three other cell lines (breast carcinoma MCF-7, HeLa S3, and melanoma A375) produced small amounts of the beta subunit of human luteinizing hormone but did not appear to secrete it. Two cell lines (the melanoma line A375 and the SV40-transformed line SV80) appeared to contain small amounts of human follicle-stimulating hormone. Sodium butyrate caused a 40-fold induction in the secretion of both HCG-alpha and HCG-beta by HeLa S3 cells, but the total amount of HCG-alpha secretion induced was 800-fold greater than that of HCG-beta. Induction was blocked by actinomycin D (1 microgram/ml) and cycloheximide (5 microgram/ml) but was not affected by 1-beta-D-arabinofuranosylcytosine at a concentration (5 microgram/ml) that blocked DNA synthesis 99%. These results indicate that a number of malignant human cell lines produce the subunits of both placental and pituitary gonadotropins and that there is frequently an excess secretion of the free alpha subunit common to these hormones.

Butyrates↗
PubMed1979

Effects of macromomycin on the ultrastructure and biological properties of cultured mammalian cells.

Macromomycin is shown to inhibit the biosynthesis of RNA, DNA, and protein in cultured cells of KB, HBL-100, SW-613, MCF-7, and A1Ab. There was no substantial increase in cell numbers in cultures containing macromomycin (5 microgram/ml), but after 24 to 48 hr the cells were two to three times the diameter of control cells with concomitant increase in cell protein. The ultrastructural changes induced by macromomycin in KB cells demonstrate an increase in nuclear size without similar changes in the size of other cytoplasmic subcellular units. It was of interest to note the general proliferation of cellular organelles and the increased occurrence of annulate lamellae followed, after prolonged treatment, by the appearance of larger numbers of lipid droplets and lysosomes; vacuoles developed to a significant extent after the cells detached from the monolayer. A1Ab cells show ultrastructural changes similar to those of KB cells when treated with macromomycin.

Antibiotics, Antineoplastic↗
PubMed1979

The immunoendocrinology of estrophilin.

Immunoglobulin from the serum of rabbits immunized with highly purified estradiol-receptor complex from calf uterine nuclei has been shown to contain specific antibodies to estrophilin by five criteria. Antibodies to calf nuclear estrophilin cross react with nuclear estradiol-receptor complexes of rat, rabbit and sheep uterus, rat endometrial and pituitary tumor, and MCF-7 human breast cancer cell line. They also react with extranuclear receptor of calf, rat, mouse, rabbit, guinea pig, monkey and sheep uterus, rat mammary, endometrial and pituitary tumor, and human breast cancer. There is no interaction of the antibody with estradiol itself. The nuclear form of estrophilin appears to bind more immunoglobulin molecules than does the cytosol form. The antibodies do not react with either the nuclear or extranuclear dihydrotestosterone-receptor complexes of rat prostate, with the extranuclear progesterone-receptor complexes of rabbit uterus, chick oviduct or rat endometrial tumor, or with rat and mouse alpha-fetoprotein. These findings indicate an immunochemical similarity among estrophilins from several mammalian species, as well as between nuclear and extranuclear forms of the receptor, but not among receptor proteins for different steroid hormones. Immunoglobulin from the serum of a goat immunized with similar antigen shows a considerably higher titer of antibodies to estrophilin. These react with nuclear and extranuclear estradiol-receptor complexes of calf uterus to produce somewhat larger entities than those formed with the rabbit antibody. Unlike the rabbit antibody, interaction with the goat antibody causes a noticeable decrease in estradiol-binding affinity of the extranuclear estrophilin as well as an apparent decrease in the total hormone-binding capacity. Specific antibodies to estrophilin offer promise as valuable reagents for receptor analysis and purification, as well as for the elucidation of many still unresolved questions concerning receptor synthesis, localization and function.

Animals↗
PubMedDate not supplied

[Estrogen dependance of the progesterone "receptor" in human tissues (author's transl)].

The concentration of specific progesterone receptor (RP) has been measured in the endometrium of 31 normal menstruating women during the two phases of their menstrual cycle, and compared with the plasma oestradiol-17 beta (E2) concentrations. There was no relationship between RP and plasma E2 when the two phases of the cycle were considered, however a correlation was observed when the follicular phase was only considered. The relationship between oestrogen and RP was then more directly studied in a breast cancer cell line in tissue culture (MCF-7). RP induction by E2 as well as oestriol was demonstrated, but oestrone was ineffective.

Endometrium↗
PubMed2026

Integrated Optimization, Genomic Characterization, and Functional Evaluation of Biogenic Selenium Nanoparticles from Bacillus licheniformis BLN313: Antibacterial and Anticancer Potential.

Microbial synthesis of selenium nanoparticles (SeNPs) offers a sustainable alternative to chemical routes, but the genetic basis of selenium handling in Bacillus remains poorly defined, which limits rational strain selection. Here, SeNP production, physicochemical characterization, and closed-genome sequencing are combined for Bacillus licheniformis BLN313. Selenite reduction peaked at 500 µg/mL Na2SeO3 (88.8% conversion; 444 ± 27 µg/mL Se0); at higher concentrations, conversion efficiency and viability diverged, indicating that tolerance and reductive capacity are distinct traits. Purified SeNPs were spherical and partially crystalline trigonal Se0 (TEM 190 ± 52 nm; DLS 166 nm, PDI 0.03; zeta potential -20.8 mV), carrying a proteinaceous capping layer confirmed by XPS, EDS, and FTIR and shown by LC-MS to be enriched in cell wall-derived metabolites. The particles were bactericidal against Micrococcus luteus (MIC 62.5 µg/mL) and Klebsiella pneumoniae (MIC 250 µg/mL) and reduced MCF-7 viability (IC50 2.7 µg/mL) while sparing MCF-10A cells. The 4.11 Mb genome (46.3% GC; ANI 99.7%, dDDH 97.8%) encodes SulP and Pit transporters, multiple trxB copies, and sulfur-metabolism and oxidative-stress genes, defining a candidate gene set for selenium uptake, reduction and detoxification. BLN313 thus provides a genetically defined platform for SeNP production in biomedical and environmental applications.

Selenium↗