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Biomedical subjects

D Dai

Publications and source records attributed to D Dai.

At least 37 records · Page 2Linked to original sources
PubMed2001

Molecular tools to reestablish progestin control of endometrial cancer cell proliferation.

OBJECTIVE: Endometrial cancers often arise in a setting of estrogen stimulation unopposed by the differentiating effects of progesterone. Our laboratory and others have previously shown that progesterone receptor down-regulation or perturbation of progesterone receptor isoform A or B expression is associated with the development of poorly differentiated endometrial cancers that are not growth inhibited by progestins. The purpose of these studies was to reestablish high progesterone receptor isoform A and B gene expressions in such endometrial cancer cells and to examine the effects of progestin treatment on cell growth and metastatic potential after this transformation. STUDY DESIGN: To induce high levels of expression of the progesterone receptor isoforms in KLE and Hec50 endometrial cancer cells, adenoviral vectors encoding the genes for progesterone receptor isoforms A and B were created. The characteristic ability of cancer cells to grow independently of anchorage to the surrounding solid matrix was measured by counting colony formation on soft agar for 8 to 14 days. Cell proliferation in response to a time course of progestin treatment was tested with flow cytometry. RESULTS: After treatment with a control vector without a progesterone receptor--encoding insert, no effect of progestin treatment on cell proliferation was found; after treatment with vectors encoding progesterone receptor isoform A or B, however, progestin treatment resulted in significant inhibition of cell growth. The anchorage-independent cell growth on soft agar assay showed that by 8 to 14 days the number of cell colonies was reduced by 50% relative to control preparations in the presence of progesterone receptor isoform A plus progestin (P <.0001, both Hec50 and KLE cell lines) and by 90% in the presence of progesterone receptor isoform B plus progestin (P <.0001, both Hec50 and KLE cell lines). Progestin treatment also resulted in a time-dependent reduction in cell proliferation as measured by flow cytometry. Although transfection with both progesterone receptor isoforms A and B reduced cell proliferation according to our assays, progesterone receptor isoform B caused a much more dramatic decrease in cell growth (P =.001, Hec50 cells; P <.0001, KLE cells). CONCLUSION: In poorly differentiated endometrial cancer cells that are resistant to progestin therapy, adenovirus-induced expressions of progesterone receptors A and B reestablish progestin control of endometrial cancer cell proliferation.

Blotting, Western↗
PubMed2001

Polymorphisms in human CYP2C8 decrease metabolism of the anticancer drug paclitaxel and arachidonic acid.

Cytochrome P450 (CYP) 2C8 is the principal enzyme responsible for the metabolism of the anti-cancer drug paclitaxel (Taxol). It is also the predominant P450 responsible for the metabolism of arachidonic acid to biologically active epoxyeicosatrienoic acids (EETs) in human liver and kidney. In this study, we describe two new CYP2C8 alleles containing coding changes: CYP2C8*2 has an Ile269Phe substitution in exon 5 and CYP2C8*3 includes both Arg139Lys and Lys399Arg amino acid substitutions in exons 3 and 8. CYP2C8*2 was found only in African-Americans, while CYP2C8*3 occurred primarily in Caucasians. Neither occurred in Asians. The frequency of the CYP2C8*2 allele was 0.18 in African-Americans, and that of CYP2C8*3 was 0.13 in Caucasians. CYP2C8*1 (wild-type), CYP2C8*2 and CYP2C8*3 cDNAs were expressed in Escherichia coli, and the ability of these enzymes to metabolize both paclitaxel and arachidonic acid was assessed. Recombinant CYP2C8*3 was defective in the metabolism of both substrates. The turnover number of CYP2C8*3 for paclitaxel was 15% of CYP2C8*1. CYP2C8*2 had a two-fold higher Km and two-fold lower intrinsic clearance for paclitaxel than CYP2C8*1. CYP2C8*3 was also markedly defective in the metabolism of arachidonic acid to 11,12- and 14,15-EET (turnover numbers 35-40% that of CYP2C8*1). Thus, CYP2C8*3 is defective in the metabolism of two important CYP2C8 substrates: the anticancer drug paclitaxel and the physiologically important compound arachidonic acid. This polymorphism has important clinical and physiological implications in individuals homozygous for this allele.

Alleles↗
PubMed2001

Assembly, annotation, and integration of UNIGENE clusters into the human genome draft.

The recent release of the first draft of the human genome provides an unprecedented opportunity to integrate human genes and their functions in a complete positional context. However, at least three significant technical hurdles remain: first, to assemble a complete and nonredundant human transcript index; second, to accurately place the individual transcript indices on the human genome; and third, to functionally annotate all human genes. Here, we report the extension of the UNIGENE database through the assembly of its sequence clusters into nonredundant sequence contigs. Each resulting consensus was aligned to the human genome draft. A unique location for each transcript within the human genome was determined by the integration of the restriction fingerprint, assembled genomic contig, and radiation hybrid (RH) maps. A total of 59,500 UNIGENE clusters were mapped on the basis of at least three independent criteria as compared with the 30,000 human genes/ESTs currently mapped in Genemap'99. Finally, the extension of the human transcript consensus in this study enabled a greater number of putative functional assignments than the 11,000 annotated entries in UNIGENE. This study reports a draft physical map with annotations for a majority of the human transcripts, called the Human Index of Nonredundant Transcripts (HINT). Such information can be immediately applied to the discovery of new genes and the identification of candidate genes for positional cloning.

Alleles↗
PubMed2001

Metabolism of chlorpyrifos by human cytochrome P450 isoforms and human, mouse, and rat liver microsomes.

One of the factors determining the toxicity of chlorpyrifos (CPS), an organophosphorus (OP) insecticide, is its biotransformation. CPS can be activated by cytochrome P450 (CYP) through a desulfuration reaction to form chlorpyrifos-oxon (CPO), a potent anticholinesterase. CPS can also be detoxified by CYP through a dearylation reaction. Using pooled human liver microsomes (HLM), a K(m(app)) of 30.2 microM and V(max(app)) of 0.4 nmol/min/mg of protein was obtained for desulfuration, and a K(m(app)) of 14.2 microM and a V(max(app)) of 0.7 nmol/min/mg of protein was obtained for dearylation. These activities are lower than those obtained from rat liver microsomes. Gender differences in humans were also observed with female HLM possessing greater activity than male HLM. Use of human CYP isoforms expressed in human lymphoblastoma cells demonstrated that CYP1A2, 2B6, 2C9*1, 2C19, and 3A4 are involved in CPS metabolism. CYP2B6 has the highest desulfuration activity, whereas dearylation activity is highest for 2C19. CYP3A4 has high activity for both dearylation and desulfuration. The use of phenotyped individual HLM demonstrated that predictions of metabolic activation and/or detoxication could be made based on relative amounts of CYP2B6, 2C19, and 3A4 in the microsomes. Thus, individuals with high CYP2C19 but low 3A4 and 2B6 are more active in dearylation than in desulfuration. Similarly, individuals possessing high levels of CYP2B6 and 3A4 have the greatest potential to form the activation product. These differences between individuals suggest that differential sensitivities to CPS may exist in the human population.

Animals↗
PubMed2001

[Expression of telomerase reverse transcriptase and its relationship with tumor suppressor gene p53 in HCC].

OBJECTIVE: To explore the expression of human telomerase reverse transcriptase (HTERT) and its relationship with tumor suppressor gene p53. METHODS: RNA-RNA in situ hybridization (ISH) technique was applied to localize the expression of HTERT mRNA and immunohistochemistry (IHC) was used to observe the p53 protein expression in 34 specimens of HCC and their corresponding non-cancerous tissues. Both the significance of HTERT mRNA expression in the different histopathologic characters and its relationship with p53 protein were investigated. RESULTS: HTERT was detected in tumor tissue in 30 of 34 cases (88.2%), and all the 6 cases of small cancer (diameter<3cm) were positive. The positive rate of HTERT mRNA was higher in the tumor tissues with low-differentiation, extra- or intra-hepatic metastasis, uncompleted fibrotic capsule and/or the non-cancerous tissue with chronic active lesions or positive HBsAg. Intensive signal was seen both in the HCC cells along the front of invasive area and in a few non-cancerous liver cells with dysplasia. Of all 34 HCC, p53 protein was presented in 25 cases. There was a correlation between the expression of HTERT mRNA and of p53 protein. The coincidence rate of the expression (HTERT+/p53+ and HTERT-/p53-) was 79.4%. More intensity of HTERT signal could be seen in the cases of positive p53. HTERT mRNA was not found in normal liver tissues. CONCLUSIONS: High expression of HTERT mRNA is proved in most cases of HCCs. HTERT expression may relate to the pathogenesis and the invasion of HCC. p53 may play a role in regulating the expression of HTERT.

Adult↗
PubMed2001

Identification of variants of CYP3A4 and characterization of their abilities to metabolize testosterone and chlorpyrifos.

CYP3A4 is the most abundant isoform of cytochrome P450 (CYP) in adult human liver. It metabolizes numerous clinically, physiologically, and toxicologically important compounds. The expression of CYP3A4 varies 40-fold in individual human livers, and metabolism of CYP3A4 substrates varies at least 10-fold in vivo. Single nucleotide polymorphisms (SNPs) in CYP3A4 were identified by direct sequencing of genomic DNA in 72 individuals from three different ethnic groups, including Caucasians, Blacks (African-Americans and African pygmies), and Asians. A total of 28 SNPs were identified, including five which produced coding changes M445T (CYP3A4*3), R162Q (CYP3A4*15), F189S (CYP3A4*17), L293P (CYP3A4*18), and P467S (CYP3A4*19). The latter four represent new alleic variants. Racial variability was observed for the frequency of individual SNPs. CYP3A R162Q was identified only in Black populations with an allelic frequency of 4%. CYP3A4 F189S and CYP3A4 M445T were identified in Caucasians with allelic frequencies 2% and 4%, respectively. L293P and P467S were only observed in Asians at allelic frequencies of 2%. The cDNAs for the F189S, L293P, M445T, and P467S mutant alleles were constructed by site-directed mutagenesis and expressed in an Escherichia coli expression system. Testosterone and the insecticide chlorpyrifos were used to assess the catalytic activities of the most common CYP3A4 allele (CYP3A4*1) and its allelic variants. CYP3A4 F189S exhibited lower turnover numbers for testosterone and chlorpyrifos, while CYP3A4 L293P had higher turnover numbers for both substrates. The turnover numbers of the CYP3A4 M445T and P467S alleles to metabolize these compounds were not significantly different from those of wild-type CYP3A4.

Alleles↗
PubMed2001

[The clinical significance of E-cadherin and alpha-catenin expression in human gastric cancer].

OBJECTIVE: To evaluate the expression of cell adhesion molecules E-cadherin (E-CD) and alpha-catenin (alpha-CA) in relation to biologic behavior of gastric cancer. METHODS: The level of E-CD and alpha-CA expression was examined in 70 cases of primary gastric cancer by immunohistochemical technique. RESULTS: The rate of E-CD and alpha-CA expression was significantly reduced in gastric cancer with more prominent malignant phenotype, such as tumor in stage III/IV with poor cell differentiation, invasion through serosa and lymph node (LN) metastasis. With the increase in the number and frequency of LN metastasis and its dissemination distal to second group LNs, the reduction in expression of the two adhesion molecules was increasingly significant. The frequency of LN metastasis and shed cancer cells in the peritoneum in patients with E-CD(-)/alpha-CA(-) and E-CD(+)/alpha CA(-) cancer were significantly higher than those in patients with E-CD(+)/alpha-CA(+) tumor. Liver metastasis was most frequent in all cases with E-CD(+)/alpha-CA(-) tumor. CONCLUSION: Down regulation of E-CD and/or alpha-CA correlates with the malignant behavior of gastric cancer. Examination of E-CD and alpha-CA expression in primary gastric cancer helps evaluate its intensity of LN metastasis, TNM staging and prognosis.

Cadherins↗
PubMed2000

Transfection of C6 glioma cells with glia maturation factor upregulates brain-derived neurotrophic factor and nerve growth factor: trophic effects and protection against ethanol toxicity in cerebellar granule cells.

Glial cells play active roles in neuronal survival, as well as neuroprotection against toxic insult. Recent studies suggest that the brain protein glia maturation factor (GMF) is involved in intracellular signaling in glia. This study investigated whether or not GMF plays a role in the survival-promoting and neuroprotective functions of glia. C6 glioma cells were transfected in vitro with GMF utilizing an adenovirus vector. The transfected cells overexpressed GMF intracellularly, but did not secrete the protein. The conditioned medium (CM) was obtained from the GMF-transfected cells (CM-GMF) and tested on primary neuronal cultures, consisting of cerebellar granule cells (CGC). The CGC cultures were utilized because these cultures have a background level of cell death, and the survival-promoting, i.e. neurotrophic effect, of the CM could be tested. In addition, since CGC cultures are ethanol-sensitive (ethanol enhances neuronal death), the neuroprotective effect of the CM against ethanol-induced cell death was tested also. We demonstrated that the CM-GMF had an enhanced neurotrophic effect as well as an increased neuroprotective effect against ethanol-induced cell death compared to control CM obtained from untransfected C6 cells (CM-Mock) or CM obtained from cells transfected with an unrelated gene (CM-LacZ). Because neurotrophins have trophic and protective effects, we investigated whether GMF-transfection upregulated the expression of neurotrophins in C6 cells. RT-PCR verified that GMF-transfected C6 cells had increased mRNA levels for BDNF and NGF. Immunoblotting corroborated the RT-PCR results and indicated that CM-GMF contained greater concentrations of BDNF and NGF protein compared to CM-Mock and CM-LacZ. A soluble TrkB-IgG fusion protein, which selectively binds BDNF and prevents its binding to the neuronal TrkB receptor, eliminated the neurotrophic effect of CM-GMF; whereas anti-NGF antibody was ineffective in preventing this effect, suggesting that the neurotrophic effect was due to BDNF. On the other hand, both the TrkB-IgG fusion protein and anti-NGF reduced neuroprotection, suggesting that BDNF and NGF both contribute to the neuroprotective effect of CM-GMF. In conclusion, GMF upregulates the expression of BDNF and NGF in C6 cells, and these factors exert neurotrophic and neuroprotective functions on primary neurons.

Adenoviridae↗
PubMed2000

Synthesis and pharmacology of a hybrid cannabinoid.

A pentacyclic hybrid cannabinoid (4) has been synthesized, which combines structural elements of traditional cannabinoids and cannabmimetic indoles. Cannabinoid 4 contains a 1-pentylindole structure fused to the 2,3-positions of the partially reduced hydroxydibenzopyran system of THC. The successful approach to 4 employed 9-benzoyl-5,7-dimethoxy-1,2,3,4-tetrahydrocarbazole (17) as the starting material. Dehydrogenation to carbazole 18, followed by demethylation and condensation with trans-p-menthadienol gave N-benzoyl hybrid cannabinoid 22, N-alkylation of which afforded target cannabinoid 4. The hybrid cannabinoid had affinity for the CB1 receptor approximately equal to that of delta8-THC (Ki = 19.3+/-3 nM), and shows comparable potency in vivo.

Animals↗
PubMed2000

Regression of the decidualized mesometrium and decidual cell apoptosis are associated with a shift in expression of Bcl2 family members.

The purpose of this study was to determine whether regression of the decidua basalis (DB), which begins on Day 14 of pregnancy in the rat, results from an intrinsic program of apoptosis regulated by Bax and Bcl2. Expression of Bax and Bcl2 and the incidence of apoptosis were evaluated throughout gestation by Western blot analysis and detection of DNA fragments. Antiprogestin (RU486) was also administered during proliferation of DB to study progesterone regulation of Bax/Bcl2 balance. Bax, the pro-apoptotic protein, was expressed at a low level throughout pregnancy, whereas Bcl2, the pro-survival partner, was most abundantly expressed on Days 8 and 10, which are a time of proliferation and decidualization, and declined to barely detectable levels thereafter. These changes resulted in a 12-fold increase in the Bax:Bcl2 ratio on Day 17 as compared with Day 8 of pregnancy (P < 0.05). DNA laddering and in situ staining of DNA fragments first became visible on Day 14 and involved 2% of cells by Days 17 and 21 (P < 0.05). Treatment with RU486 on Day 9 enhanced Bax and suppressed Bcl2 within 6 h, increasing the Bax:Bcl2 ratio sixfold (P < 0.05). Apoptosis was minimal at 6 h and increased to 9% of cells by 24 h (P < 0.05). Thus, progesterone appears to regulate the apoptotic threshold of stromal cells by modulating Bax and Bcl2 expression.

Animals↗
PubMed2000

Performance of a 62Zn/62Cu generator in clinical trials of PET perfusion agent 62Cu-PTSM.

UNLABELLED: The 62Zn/62Cu PET generator can be inexpensively produced and distributed from a single production site operating under typical good manufacturing practice guidelines. It therefore has the potential to greatly facilitate development of clinically practical PET. We report generator performance in a study in which 62Cu-pyruvaldehyde-bis(n4-methylthiosemicarbazone (PTSM) myocardial perfusion imaging is compared with 99mTc-sestamibi in the diagnosis of coronary artery disease. The 62Zn/62Cu generator is an improved version of a previously reported system that employs automated synthesis of 62Cu-PTSM. With this approach, the cumbersome step of 18C purification has been eliminated. METHODS: The 62Zn (9.3 h half-life) parent isotope is prepared by proton bombardment of natural copper at 33 MeV. A typical target irradiated with 37.5 microA/h is delivered by 12:00 PM on the day it is to be processed. Purified 62Zn obtained from the target is loaded onto the generator column in 2 mol/L HCl. The generator is eluted using an internal three-channel peristaltic pump, which delivers 2.25 mL eluant (1.8 mol/L NaCl, 0.2 mol/L HCl) through the generator column to elute the 62Cu in 40 s. The same pump simultaneously pumps an equal volume of buffer (0.4 mol/L NaOAc) and 1 mL ligand solution (2 ppm PTSM, 2% EtOH) passing it through a septum into a 35-cc syringe preloaded with 28 mL sterile water. This solution is thoroughly mixed by agitation of the syringe and injected as a bolus through a 0.2 microm filter. The generator is eluted twice before shipping, providing quality assurance samples, and shipped to the clinical site by overnight delivery. Complete quality assurance testing is performed the evening before the generator reaches the clinical site. RESULTS: A total of 34 generators have been produced and shipped to 2 clinical sites for a phase III Food and Drug Administration study. The load activity on the generators at 8:00 AM the day of clinical use was 1.7+/-0.2 GBq (46.7+/-5.6 mCi), and yield was 72%+/-16%. Breakthrough of 62Zn was undetectable by high-purity germanium spectroscopy for all units. Radiochemical purity was 95.4%+/-2.4%. Volume delivered, pH, sterility, and bacterial endotoxin tests yielded passing results on all generators. The entire process of generator production, from target receipt to generator shipment, took less than 6 h and cost approximately $1000, including shipping charges and cyclotron cost. A total of 68 patients were injected with 2 62Cu-PTSM doses, with a mean injected activity of 0.8+/-0.2 GBq (20.5+/-5.3 mCi) with no adverse side effects. CONCLUSION: Results of this work confirm that the 62Zn/62Cu generator is an easily produced, transportable, and inexpensive source of PET radiopharmaceuticals, which can expand the field of clinical PET imaging by providing radiopharmaceuticals to sites not associated with cyclotrons.

Clinical Trials, Phase III as Topic↗
PubMed2000

[The causes of death in patients with malignant biliary obstruction treated by metallic stent and/or biliary drainage].

OBJECTIVE: To analyze the causes of death in patients with malignant bile duct obstruction treated by metallic stent endoprosthesis and biliary drainage. METHODS: One hundred and seven patients with malignant bile duct obstruction were so treated. Of the 107 patients, 12 (11.2%) died within 30 days after operation. The bile duct obstruction in the 12 cases was due to primary hepatic cancer(n = 4), metastasis of lymph nodes in hepatic hilus(n = 3) and bile duct cancer(n = 5). RESULTS: Five patients died of biliary infections, 4 patients died of hepatic coma, and 2 patients died of cancerous cachexia and 1 patient died of biliary tract hemorrhage. CONCLUSION: The major causes of postoperative death of patients with biliary obstruction treated by metallic stent to support bile duct are biliary infection and hepatic encephalopathy.

Aged↗
PubMed2000

[Clinical and pathological characteristics of gastric stump cancer and recurrent gastric stump cancer].

OBJECTIVE: To study early diagnosis and rational treatment of gastric stump cancer and recurrent gastric stump cancer. METHODS: The data of 27 patients with gastric stump cancer and 23 patients with recurrent gastric stump cancer were analysed retrospectively during the past 20 years based on their clinical and pathological findings. RESULTS: The onset time was 10 years or more for gastric stump cancer and less than 10 years for recurrent gastric stump cancer after gastrectomy. Gastroscopy was better than double contrast radiography in diagnosis. Partial resection of gastric stump was performed for patients with early cancer at the anastomotic stoma of gastrojejunostomy. Total resection of gastric stump was made for other patients. Lymph node of the mesojejunum were dissected routinely because their metastasis rate was about 40.0% in patients with cancer of anastomotic stoma. The effect of surgical treatment was poor for patients with advanced cancer. CONCLUSIONS: Regular gastroscopy is useful in detecting gastric stump cancer in patients with gastric cancer after gastrectomy, or in those years after removal benign. Malignancy was more evident in recurrent gastric stump cancer than in primary gastric cancer. Prognosis is good for early stage cases and poor for late stage cases after surgical treatment.

Adult↗
PubMed2000

[A case-control study on the risk factors of esophageal cancer in Linzhou].

OBJECTIVE: To explore the characteristics of prevalence and influencing factors on the genesis of esophageal cancer. METHODS: A population-based 1:1 matched case-control study was conducted in Linzhou. A total number of 352 pairs of cases and controls matched on sex, age and neighborhoods. Data was analysed by SAS software to calculate the odds ratio of and to evaluate the relative risks. RESULTS: It was found that lower socio-economic status, environmental pollution around the residential areas, lampblack in room, lower body mass index (BMI), more pickled food intake, cigarette smoking, alcoholic drinking, vigor mental-trauma and depression were risk factors of esophageal cancer. It also showed that the subjects having had history of upper digestive tract operation, dysplasia of esophagus and family history of carcinoma markedly increased the risks of developing esophageal cancer. CONCLUSION: Esophageal cancer seemed to be resulted from the combination of genetic and environmental factor, hence called for of medical surveillance and comprehensive prevention.

Adult↗
PubMed1999

Ventrally emigrating neural tube cells differentiate into heart muscle.

A population of ventrally emigrating neural tube cells has been shown to migrate along the vagus nerve and contribute to the development of the gastrointestinal tract. Since the vagus also goes to the heart, we sought to determine if these cells migrated into the heart. Neural tube cells were tagged with replication-deficient retroviral vectors containing the LacZ gene, to permanently label their progeny. The virus was microinjected into the lumen of the caudal hindbrain of chick embryos on day 2. Embryos were later processed for the detection of LacZ positive cells. Labeled cells were initially confined to the neural tube. Later, they migrated in association with the vagus nerve into the heart, where they were located in the myocardium. Labeled cells were identified as cardiac muscle cells of non-neural crest origin, with specific markers. It is concluded that some cardiac muscle cells differentiate from the neural tube cells.

Animals↗
PubMed1999

Ventrally emigrating neural tube cells contribute to the formation of Meckel's and quadrate cartilage.

A population of multipotential neuroepithelial cells originating in the ventral portion of the hindbrain neural tube has been shown recently to emigrate at the site of attachment of the trigeminal nerve. These ventrally emigrating neural tube cells populate the mesenchyme of the first pharyngeal (branchial) arch. Because the Meckel's and the quadrate cartilage develop from this mesenchyme, we sought to determine whether these ventrally emigrating neural tube cells contributed to their development. The ventral neural tube cells were tagged with a replication-deficient retroviral vector containing the LacZ gene. This method permanently labels the descendents of the neural tube cells; thus, they can be subsequently tracked during development. The viral concentrate was microinjected into the lumen of the rostral hindbrain of chick embryos, after the emigration of neural crest is finished, on embryonic day 2 (stage 14). In control embryos, the virus was placed on top of the neural tube. Embryos were killed on days 3, 4, and 7 and processed for the detection of LacZ-positive cells. By day 7, the Meckel's and the quadrate cartilage can be easily recognized. LacZ-positive cells were seen in both cartilages. They were located in perichondrium and in the cartilage. Immunostaining with the neural crest cell marker HNK-1 indicated that the LacZ-positive cells were HNK-1 negative. The HNK-1-positive neural crest-derived cells were located in the cartilage but not in the perichondrium. These results indicate that the chondrocytes in the Meckel's and the quadrate cartilage differentiate from two sources of cells; the ventrally emigrating neural tube cells and the neural crest. The developmental significance of differentiation of cartilage from the ventral neural tube cells and of the heterogeneous origin of chondrocytes in morphogenesis remains to be established. Dev Dyn 1999;216:37-44.

Animals↗
PubMed1999

Progesterone-regulated determinants of stromal cell survival and death in uterine decidua are linked to protein kinase C activity.

This study examined the role of protein kinase C enzymatic activity as a physiologic determinant of stromal cell death in decidua basalis (DB) during pregnancy. The expression of epidermal growth factor receptor (EGF-R) and Bcl2 was used as an indicator of stromal cell proliferation/survival, whereas Bax and the occurrence of apoptosis provided an index of cell death. Stromal cell cycle progression during pregnancy and after in vivo administration of phorbol esters was analyzed by flow cytometry. DB were isolated from pregnant rats between Days 8 and 21 of pregnancy and prepared for immunohistochemistry, Western blotting procedures, or flow cytometry. The results showed that stromal cells were actively proliferating on Days 8 and 10, whereas the frequency of cell death by apoptosis increased progressively between Days 14 and 21 (Day 22 is term). The proliferative stage was characterized by low PKC activity and high levels of EGF-R and Bcl2 expression. On the other hand, DB regression (Days 14-21) was marked by an elevation in endogenous PKC activity and Bax expression; EGF-R and Bcl2 were suppressed. Administration of phorbol 12-myristate, 13-acetate (0.4 micromole/kg) induced apoptosis on Day 10. Additionally, antiprogestin (RU-486) given on Day 9 induced PKC activity and Bax expression within 6 h and suppressed Bcl2 and EGF-R. By 12 h, RU-486 enhanced percent apoptotic cells. Thus, enhanced levels of PKC activity were closely linked to stromal cell apoptosis.

Animals↗