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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 434 (1999), S. 23-28 
    ISSN: 1432-2307
    Keywords: Key words p16 ; Rb ; Uterus ; Endometrium ; MTS1 ; Carcinoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  P16MTS1/CDKN1 and the retinoblastoma protein Rb are both involved in negative regulation of G1/S progression in the mammalian cell cycle. Inactivation of one of these tumour suppressor genes is involved in many malignant tumours, and in some studies a negative correlation of p16 and Rb expression has been found. In order to study this interaction in endometrial carcinogenesis, we investigated 36 endometrial carcinomas, 11 cases of hyperplasia, 23 normal endometrial samples, and two uterine carcinoma cell lines by immunohistochemistry or RT-PCR. Rb was expressed in normal endometrial epithelium, hyperplasia, cell lines, and most carcinomas; negative immunostaining was only detected in 1 of 36 tumours. In contrast, p16 expression was weak in normal endometrium and increased in most cases of hyperplasia, but negative or minimally positive in 74% of the carcinomas and the Hec1B adenocarcinoma cell line, and there was no significant association with Rb immunostaining. Strikingly high p16 expression was found in foci of squamous metaplasia within hyperplastic or carcinomatous tissue. Deletion and mutation analysis of the p16 gene was performed in DNA from microdissected tumour samples and cell lines. No p16 deletion was found, and mutations were detected in only one tumour sample and Skut1B uterine mixed mesodermal tumour cells. Our data indicate that in spite of low or absent p16 expression, genetic alterations of the p16 and Rb tumour suppressor genes are rare in endometrial carcinogenesis.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-7217
    Keywords: cytokines ; leukemia inhibitory factor ; estrogen regulation ; progestin regulation ; promoter ; MDA-MB-231 cells ; T47D cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Leukemia inhibitory factor (LIF) is a pleiotropic inflammatory cytokine. A potential role for LIF in the pathogenesis of human breast cancer was recently indicated by the finding that LIF is produced by MDA-MB 231 breast cancer cells and that it stimulates proliferation of the T47D and MCF-7 breast cancer cell lines. Despite its role as a possible therapeutic target in breast cancer, the transcriptional regulation of the LIF gene in breast cancer cells has not been investigated so far. In this context, we investigated the regulation of the human LIF promoter (human LIF666-luciferase) by ovarian steroids in transient transfection assays in MDA-MB 231 and T47D cells. Since the MDA-MB 231 cells are devoid of both estrogen (ER) and progesterone (PR) receptors, these cells were co-transfected with the respective receptor expression vector. Estradiol induced no stimulation in either T47D or ER-transfected MDA-MB 231 cells. Treatment with the progesterone agonist MPA (medroxy-progesterone acetate) resulted in induction of LIF transcription in PR-transfectant MDA-MB 231 cells, while it had no effect in T47D cells. Both PR isoforms (PR-B and PR-A) were effective in inducing the LIF promoter in MDA-MB 231 cells, and this effect was inhibited by the progestin antagonist RU 486. The stimulatory effect of MPA was maintained on deletion constructs (h274LIF-Luc, h148LIF-Luc and h82LIF-Luc), indicating that 82 bp are sufficient to mediate this effect. Our results indicate that the LIF promoter is transcriptionally active in human breast cancer cells and its activity can be modulated by progestins and anti-progestins in cells expressing the LIF protein, which might have therapeutic implications.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-7217
    Keywords: breast cancer ; cyclin D1 ; MPA ; proliferation ; T47D cell line
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract While progesterone is a known differentiation-inducing factor in the human endometrium, for the breast epithelium both proliferation-inducing and -inhibiting effects have been described. Cyclin D1, which is required for cell cycle progression in G1 and has been shown to play an important role in the pathogenesis of breast cancer has been implicated as a possible mediator of such effects. In the present study we thus investigated the effects of the progestin agonist MPA (medroxy-progesterone acetate) on proliferation of T47D breast cancer cells. In parallel experiments, the regulation of the human cyclin D1 promoter as well as cyclin D1 protein levels under the influence of MPA were studied. Our results show an increase of proliferative activity in T47D cells after 24 and 48 h of MPA treatment follwed by inhibition of proliferation after 72 h. In Western blot analysis an increased expression level of cyclin D1 protein can be observed after 24 h of MPA stimulation, while at 72 h the protein levels are barely detectable. Transient transfection experiments with a luciferase reporter plasmid containing the human cyclin D1 promoter showed an induction of the promoter after 24 and 36 h of MPA treatment followed by a reduction in promoter activity. In conclusion, our results confirm the existence of a biphasic response of T47D cell proliferation in response to MPA treatment, consisting of stimulation of proliferation followed by inhibition, and further implicate cyclin D1 as a mediator of these effects, since the cyclin D1 promoter shows a similar biphasic response in this context.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Endometrium ; Molekulare Endokrinologie ; Vaginale Sonographie ; Substitutionsbehandlung ; Tamoxifen ; Key words Endometrium ; Molecular endocrinology ; Vaginal sonography ; Hormone replacement therapy ; Tamoxifen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The ovarian steroid hormones estradiol and progesterone exert their effect by interacting with their intracellular receptors, which, after ligand binding translocate to the nucleus and bind to the promoter regions of target genes. The consequence is a change in the transcription rate of the target genes, followed by a change in production of the corresponding proteins. Target genes of the sexual steroid hormoness include cytokines and growth factors, among them CSF-1, TGF-β and LIF. The rhythm and activity of steroidogenesis, receptor modulation and transcription are reflected by cycle-specific proliferation and differentiation processes in the endometrium. Quantitative and/or qualitative molecular endocrinology is of increasing interest for better definition of morphological changes, although, as yet, the pathological laboratory test is of much less practical consequence than a suspicious vaginal sonography. In spite of the high standard of ultrasound techniques, however, most cases with slightly increased endometrial thickness show histologically benign changes of the endometrium rather than endometrial precancer or cancer. This is especially true for perimenopausal women with no other clinical findings. Yet, the cancer risk is increased in women under tamoxifen therapy. Hence, as a rule, these cases, when endometrial thickness exceeds 5 mm, need a diagnostic biopsy or abrasio.
    Notes: Zusammenfassung Die Sexualhormone Östradiol und Progesteron wirken auf molekularer Ebene über intrazelluläre Rezeptoren, die nach Bindung ihrer Liganden in den Zellkern translozieren und dort an die Promotorregion verschiedener Zielgene binden. Die Folge ist eine veränderte Transkriptionsrate und damit eine veränderte Produktion der entsprechenden Proteine. Zu den Zielgenen der Sexualhormone gehören u.a. Zytokine und Wachstumsfaktoren wie z.B. TGF-β, IL-1, CSF-1 und LIF. Die zeitliche Gangart und Aktivität der Steroidogenese, Rezeptormodulation und Transkription spiegeln sich morphologisch in den bekannten Proliferations- und Differenzierungsvorgängen im Endometrium wider. Umgekehrt gehen diese und deren Störungen mit biochemisch faßbaren, quantitativen und/oder qualitativen Veränderungen einher. Noch ist aber der endokrinologische Laborbefund weit weniger als die Vaginalsonographie Ausgangspunkt der Entscheidung zur Endometriumbiopsie oder Abrasio. Große Bedeutung kommt diesem Zusammenhang in der Perimenopause zu. In ≥90% der Frauen, die wegen des sonographischen Verdachts auf ein Endometriumkarzinom abradiert werden, läßt sich morphologisch nur ein „klimakterisches Übergangsendometrium” feststellen. Eine Ausnahmesituation besteht allerdings bei Frauen unter Behandlung mit Tamoxifen. Wegen des erhöhten Karzinomrisikos sollten diese ab einer sonographischen Endometriumdicke von ≥5 mm grundsätzlich histologisch untersucht werden.
    Type of Medium: Electronic Resource
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