65 research outputs found
Progestogenic effects of tibolone on human endometrial cancer cells
Tibolone, a synthetic steroid acting in a tissue-specific manner and used
in hormone replacement therapy, is converted into three active
metabolites: a Delta(4) isomer (exerting progestogenic and androgenic
effects) and two hydroxy metabolites, 3 alpha-hydroxytibolone (3
alpha-OH-tibolone) and 3beta-OH-tibolone (exerting estrogenic effects). In
the present study an endometrial carcinoma cell line (Ishikawa PRAB-36)
was used to investigate the progestogenic properties of tibolone and its
metabolites. This cell line contains progesterone receptors A and B, but
lacks estrogen and androgen receptors. When tibolone was added to the
cells, complete conversion into the progestogenic/androgenic Delta(4)
isomer was observed within 6 d. Furthermore, when cells were cultured with
tibolone or when the Delta(4) isomer or the established progestagen
medroxyprogesterone acetate was added to the medium, marked inhibition of
growth was observed. Interestingly, 3 beta-OH-tibolone also induces some
inhibition of growth. These growth inhibitions were not observed in
progesterone receptor-negative parental Ishikawa cells, and
progestagen-induced growth inhibition of PRAB-36 cells could readily be
reversed using the antiprogestagen Org-31489. Upon measuring the
expression of two progesterone-regulated genes (fibronectin and
IGF-binding protein-3), tibolone, the Delta(4) isomer and
medroxyprogesterone acetate showed similar gene expression regulation.
These results indicate that tibolone, the Delta(4) metabolite, and to some
extent 3 beta-OH-tibolone exert progestogenic effects. Tibolone and most
likely 3 beta-OH-tibolone are converted into the Delta(4) metabolite
Consequences of loss of progesterone receptor expression in development of invasive endometrial cancer
PURPOSE: In endometrial cancer, loss of progesterone receptors (PR) is
associated with more advanced disease. This study aimed to investigate the
mechanism of action of progesterone and the loss of its receptors (PRA and
PRB) in development of endometrial cancer. EXPERIMENTAL DESIGN: A
9600-cDNA microarray analysis was performed to study regulation of gene
expression in the human endometrial cancer subcell line Ishikawa PRAB-36
by the progestagen medroxy progesterone acetate (MPA). Five MPA-regulated
genes were selected for additional investigation. Expression of these
genes was studied by Northern blot and by immunohistochemistry in Ishikawa
subcell lines expressing different PR isoforms. Additionally, endometrial
cancer tissue samples were immunohistochemically stained to study the in
vivo protein expression of the selected genes. RESULTS: In the PRAB-36
cell line, MPA was found to regulate the expression of a number of
invasion- and metastasis-related genes. On additional investigation of
five of these genes (CD44, CSPG/Versican, Tenascin-C, Fibronectin-1, and
Integrin-beta 1), it was observed that expression and progesterone
regulation of expression of these genes varied in subcell lines expressing
different PR isoforms. Furthermore, in advanced endometrial cancer, it was
shown that loss of expression of both PR and E-cadherin was associated
with increased expression CD44 and CSPG/Versican. CONCLUSION: The present
study shows that progestagens exert a modulatory effect on the expression
of genes involved in tumor cell invasion. As a consequence, loss of PR
expression in human endometrial cancer may lead to development of a more
invasive phenotype of the respective tumor
Improved canine exome designs, featuring ncRNAs and increased coverage of protein coding genes OPEN
By limiting sequencing to those sequences transcribed as mRNA, whole exome sequencing is a costefficient technique often used in disease-association studies. We developed two target enrichment designs based on the recently released annotation of the canine genome: the exome-plus design and the exome-CDS design. The exome-plus design combines the exons of the CanFam 3.1 Ensembl annotation, more recently discovered protein-coding exons and a variety of non-coding RNA regions (microRNAs, long non-coding RNAs and antisense transcripts), leading to a total size of ≈152 Mb. The exome-CDS was designed as a subset of the exome-plus by omitting all 3' and 5' untranslated regions. This reduced the size of the exome-CDS to ≈71 Mb. To test the capturing performance, four exome-plus captures were sequenced on a NextSeq 500 with each capture containing four precapture pooled, barcoded samples. At an average sequencing depth of 68.3x, 80% of the regions and well over 90% of the targeted base pairs were completely covered at least 5 times with high reproducibility. Based on the performance of the exome-plus, we estimated the performance of the exome-CDS. Overall, these designs provide flexible solutions for a variety of research questions and are likely to be reliable tools in disease studies. In 2014, the first report detailing the design and performance of a whole exome sequencing (WES) enrichment assay for the dog was published by our group 1 . Aiming to selectively sequence all the regions that are transcribed to mRNA, WES is a reliable tool used to identify disease-causing or predisposing mutations at a fraction of the price of whole genome sequencing (WGS) studies. A limitation of WES is that it is based on our current knowledge of the annotation of the genome and that many disease causing mutations are likely to fall outside protein-coding regions. With new information becoming available, updates and extensions are required. Recently, an improved annotation for the dog genome has been published and new data on non-protein coding genes has been obtained 2 . Based on this data, two new target enrichment designs for dogs, called the exome-plus and the exome-CDS, were developed. The exome-plus offers the most comprehensive design. The exome-CDS is a subset of the exome-plus, focusing on the coding DNA sequences (CDS) by excluding the 3′ and 5′ untranslated regions (UTRs). Thes
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