Successful management by recombinant activated factor VII in a case of disseminated intravascular coagulopathy caused by obstetric hemorrhage.

Postpartum hemorrhage (PPH) is a life-threatening emergency in obstetrics. Although recombinant activated factor VII (rFVIIa) has become used for the treatment of some cases of massive hemorrhage, its applications in the field of obstetrics are still limited. We describe a case of successful treatment with rFVIIa for PPH due to placenta accreta. The patient was a 33-year-old woman with placental previa. Cesarean section (CS) was performed at gestational week 35. During CS, there was massive hemorrhage due to placenta accreta. After CS, disseminated intravascular coagulopathy and hypovolemic shock were diagnosed. The PPH was not controlled by transfusion therapy. On the fourth day after CS, rFVIIa (90 microg/kg x 2) was given because of the persistent PPH. Bleeding decreased and no further transfusion was required from 2 days after administration. rFVIIa was useful in the treatment of this case of obstetric hemorrhage.

Expression of stathmin in human uterus and decidualizing endometrial stromal cells.

The cytosolic phosphoprotein stathmin is upregulated at the site of embryo implantation in the rodents. However, stathmin expression in the human uterus has not yet been investigated. The distribution of uterine and placental stathmin was analyzed by immunohistochemistry, while stathmin mRNA expression was detected in endometrial tissues by the reverse transcriptase-PCR. Cultured endometrial stromal cells were used to investigate whether stathmin plays a role in decidualization. Stathmin is expressed specifically in the glandular epithelium and the stromal cells of human endometrial tissue. It is also expressed by cytotrophoblasts and extravillous trophoblasts, but not by syncytiotrophoblasts or decidual tissues during the first trimester of pregnancy. When stromal cells isolated from normal endometrial tissues were cultured and stimulated to decidualize by progesterone (P4) plus estrogen or dibutyryl cyclic 3',5'-AMP, their total and phosphorylated stathmin levels decreased. Knocking down stathmin expression in the cultured stromal cells using small interfering RNA, before the cells were exposed to the decidualizing agents, significantly suppressed decidualization, as indicated by the decreased expression of IGF-binding protein-1 and prolactin. Stathmin is differently expressed in human endometrial and placental cells and may participate in the decidualization of endometrial stromal cells.

Hypoxia-inducible factor-1 transactivates transforming growth factor-beta3 in trophoblast.

Hypoxia occurs during the development of placenta in the first trimester and is implicated in trophoblast differentiation. Intervillous blood flow increases after 10 wk of gestation and results in exposure of trophoblast cells to oxygen. Before this time, low oxygen appears to prevent trophoblast differentiation toward an invasive phenotype. The oxygen-regulated early events of trophoblast differentiation are mediated by TGF-beta3. TGF-beta3 plays a vital role in trophoblast differentiation, and its overexpression can be found in preeclamptic placenta. We sought to determine the mechanism of TGF-beta3 expression through hypoxia-inducible factor (HIF)-1. We show that HIF-1alpha and TGF-beta3 are overexpressed in preeclamptic placenta. Hypoxia not only transactivates the TGF-beta3 promoter activity but also enhances endogenous TGF-beta3 expression. Using the TGF-beta3 promoter deletion mutants, we show that the region between -90 and -60, which contains a putative HIF-1 consensus motif, is crucial for HIF-1-mediated transactivation. Electrophoretic mobility shift assays show that HIF-1 binds to the oligonucleotide containing the HIF-1 motif. Also, introduction of an antisense oligonucleotide for HIF-1 diminishes TGF-beta3 expression during hypoxia, indicating that the up-regulation of TGF-beta3 by hypoxia is mediated through HIF-1. Our results provide evidence that regulation of TGF-beta3 promoter activity by HIF-1 represents a mechanism for trophoblast differentiation during hypoxia.

Establishment of a HPV and p53-mutation-negative human cell line (CA) derived from a squamous carcinoma of the uterine cervix.

OBJECTIVE: Well-characterized human cancer cell lines are important research resources for studying cancer cell biology, as well as for developing new strategies against cancer cell growth and progression. We present a new cell line, CA, established from an invasive non-keratinizing squamous cell carcinoma of the uterine cervix in 36-year-old patient. METHODS: We measured the doubling time of CA cells. To investigate the tumorigenicity of CA, cells were inoculated subcutaneously into the back of nude mice. Several tumor markers were analyzed using culture media by EIA. PCR-based analyses were performed to examine the human papillomavirus (HPV) status and telomerase activity. CA was also screened for p53 mutation using the sequencing technique. RESULTS: The cells show rapid growth in culture with a doubling time of 14.3 h and high migration activity. Monolayer-cultured cells were polygonal, showing a pavement-like arrangement and a tendency to pile up without contact inhibition. Subcutaneous transplantation of the CA cells into nude mice formed solid tumors that were histologically diagnosed as squamous cell carcinoma, whereas no metastasis was observed. Cultured CA cells produced SCC, CEA, TPA, CA125 and SLX. Genetic and molecular analyses revealed high telomerase activity and the absence of HPV DNA. No p53 mutation was observed in this cell line. CONCLUSION: These properties suggest that CA is an aggressive cervical carcinoma cell line and may serve as a useful experimental model for studying HPV role in cervical carcinogenesis.

A novel phenoxazine derivative suppresses proliferation of human endometrial adenocarcinoma cell lines, inducing G2M accumulation and apoptosis.

We examined the effects of a novel phenoxazine, 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one (Phx), which was produced by the reaction of 2-amino-5-methyl-phenol with bovine hemoglobin on the proliferation of human endometrial adenocarcinoma cell lines, EN and KLE cells, and on induction of apoptosis and G2M arrest in these cells. Phx inhibited proliferation of these cell lines in a dose- and time-dependent manner, i.e., the inhibition rate of proliferation of EN and KLE cells was 43% and 40%, respectively, in the presence of 50 micro M Phx, and 75% and nearly 100%, in the presence of 100 micro M Phx, after 2 days. When these endometrial adenocarcinoma cells were incubated with a medium containing 100 micro M Phx for 24 h, accumulation of EN and KLE cells in the S and G2M phase and that of apoptotic cells were demonstrated by flow cytometry. Apoptosis of these cells caused by Phx was unlikely to be associated with p53, Bax, and Bcl-2, because the levels of these proteins were not altered regardless of the presence or absence of Phx. The present results suggest that Phx demonstrates antitumor activity against human endometrial adenocarcinoma cell lines EN and KLE cells, by inducing both cell cycle accumulation at S and G2M and apoptosis associated with p53, Bcl-2 and Bax insensitive pathways.

Establishment of a new human cell line (EN) with TP53 mutation derived from endometrial carcinoma.

We present a new cell line, EN, established from an invasive endometrioid adenocarcinoma of the uterine corpus in 50-year-old patient. The cells show rapid growth in culture with a doubling time of 24.4 hours and high migration activity. Monolayer-cultured cells were polygonal in shape and showed a tendency to pile up without contact inhibition. Subcutaneous transplantation of the EN cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma, whereas no metastasis was observed. Cultured EN cells produced tissue polypeptide antigen. Genetic and molecular analyses revealed high telomerase activity and estrogen receptor beta but not alpha expression. Using the polymerase chain reaction-single strand conformation polymorphism technique, we have screened EN cells for TP53 mutation in exons 5-8. A mobility shift was observed in this cell line in exon 8. A nucleotide insertion (CGT-->CAGT) was detected at codon 273, which resulted in a creation of a stop codon at codon 308. This cell line thus appears to represent the development of a more malignant clone with divergent receptor function and growth behavior, and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium.

Establishment and characterization of a new human cell line (EJ) derived from endometrial carcinoma.

We present a new cell line, EJ established from an invasive endometrioid adenocarcinoma of the uterine corpus in a 56-year-old patient. The cells show rapid growth in culture with a doubling time of 16 h and high migration activity. Monolayer-cultured cells were polygonal in shape showing a tendency to pile up without contact inhibition. Subcutaneous transplantation of the EJ cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma, whereas no metastasis was observed. Cultured EJ cells produced tissue polypeptide antigen (IPA). Genetic and molecular analyses revealed high telomerase activity but not estrogen receptor alpha expression. Using the DNA sequencing technique, we have screened EJ cells for p53 mutation in exon 5 to 8 but no mutation of p53 was observed. This cell line appears to represent the development of a more malignant clone with divergent receptor function and growth behavior, and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium.