[show abstract][hide abstract] ABSTRACT: This study was performed to compare the expression of CD44 in endometrial stromal cells (ESCs) of women with and without endometriosis and to evaluate the role of CD44 in the adherence of ESCs to peritoneal mesothelial cells (PMCs).
A PMC adherence assay was performed to evaluate the adherence of ESCs to PMCs in women with and without endometriosis. The expression of CD44 mRNA was measured by reverse transcription-polymerase chain reaction. CD44 protein was evaluated by Western blot analysis.
There were no significant differences in the expression of CD44 mRNA and protein in ESCs or in the binding of ESCs to PMCs between patients with endometriosis and controls. Although the expression of CD44 protein was decreased in both women with endometriosis and controls after anti-CD44 antibody treatment, there was no effect on binding of ESCs to PMCs. Treatment of ESCs with peritoneal fluid from endometriosis patients resulted in a significant increase in binding of ESCs to PMCs compared to untreated ESCs in the endometriosis group.
This study demonstrates that the expression of CD44 protein in ESCs from women with endometriosis might not be directly associated with adherence to PMCs.
[show abstract][hide abstract] ABSTRACT: This study examined the molecular mechanisms of apicidin in the modulation of human ovarian cancer SKOV-3 cells invasion and migration. Apicidin markedly decreased histone deacetylase 4 (HDAC4) expression and blocked cell migration and invasion. Cell migration was inhibited via down-regulation of matrix metalloproteinase-2 (MMP-2) and up-regulation of RECK in the HDAC4-blocked SKOV-3 cells. Apicidin significantly suppressed the binding of HDAC4 to Sp1 binding elements of the RECK promoter via repression of HDAC4. In an in vivo model, apicidin suppressed the growth of transplanted SKOV-3 cells by down-regulating HDAC4 and MMP-2. Apicidin may potentially be used as an anti-cancer agent for inhibition of cancer cell migration and invasion through the repression of MMP-2 which is related to the reduction of HDAC4.
Cancer letters 07/2012; 325(2):189-99. · 4.86 Impact Factor
[show abstract][hide abstract] ABSTRACT: Histone deacetylase (HDAC) inhibitors are a promising new class of anticancer agents that act by inhibiting cell proliferation and inducing apoptosis in a variety of cancer cells. Although apicidin acts as a potent HDAC inhibitor, the precise mechanism for its anti-tumor activity in human endometrial cancer cells is not completely understood. This study examined the anti-tumor effects of apicidin in Ishikawa cancer cells. The level of cell proliferation, the stage of the cell cycle, and apoptosis were measured after the apicidin treatment. Apicidin significantly inhibited the proliferation of Ishikawa cells in a dose-dependent manner. In addition, apicidin markedly up-regulated the p21(WAF1) and down-regulated the expression of cyclins (A, B1, D1, or E), and CDKs (2 or 4), which leading to cell cycle arrest. Cell cycle analysis showed that the apicidin treatment increased the proportion of cells in the G1 phase, and decreased the ratio of cells in the S phase in a dose-dependent manner. Apicidin significantly increased the sub-G1 population and the number of TUNEL positive apoptotic cells compared with the untreated control. These results were confirmed by poly-ADP ribose polymerase (PARP), an 85-kDa fragment resulting from PARP cleavage, where apicidin increased the level of PARP cleavage and caspase-3 activity in 1.0 microM apicidin-treated cells. Apicidin-induced apoptosis through caspase-3 activation was confirmed by the increase in the release of cytochrome c and the decrease in the Bax/Bcl-2 ratio. These results suggest that apicidin has anti-tumor properties on endometrial cancer cells by inducing selectively the genes related to cell cycle arrest and apoptosis.
[show abstract][hide abstract] ABSTRACT: In many clinical situations which cause thymic involution and thereby result in immune deficiency, T cells are the most often affected, leading to a prolonged deficiency of T cells. Since only the thymic-dependent T cell production pathway secures stable regeneration of fully mature T cells, seeking strategies to enhance thymic regeneration should be a key step in developing therapeutic methods for the treatment of these significant clinical problems. This study clearly shows that receptor activator of NF-kappaB ligand (RANKL) stimulates mouse thymic epithelial cell activities including cell proliferation, thymocyte adhesion to thymic epithelial cells, and the expression of cell death regulatory genes favoring cell survival, cell adhesion molecules such as ICAM-1 and VCAM-1, and thymopoietic factors including IL-7. Importantly, RANKL exhibited a significant capability to facilitate thymic regeneration in mice. In addition, this study demonstrates that RANKL acts directly on the thymus to activate thymus regeneration regardless of its potential influences on thymic regeneration through an indirect or systemic effect. In light of this, the present study provides a greater insight into the development of novel therapeutic strategies for effective thymus repopulation using RANKL in the design of therapies for many clinical conditions in which immune reconstitution is required.
Experimental and Molecular Medicine 03/2008; 40(1):59-70. · 2.57 Impact Factor
[show abstract][hide abstract] ABSTRACT: Histone deacetylase (HDAC) inhibitors are promising new class of anticancer agents that act by inhibiting cell proliferation and inducing cell cycle arrest of various cancer cells. Psammaplin A (PsA) is a phenolic natural product that has been isolated from marine sponges, and has been suggested to be a promising novel HDAC inhibitor. However, the precise mechanism of PsA as a HDAC inhibitor is poorly understood. This study investigated the anti-tumor effect of PsA on endometrial human cancer cells.
The cell proliferation, cell cycle, and apoptosis were measured in Ishikawa endometrial cancer cells after PsA treatment.
PsA significantly inhibited the proliferation of Ishikawa cells in a dose-dependent manner. PsA markedly induced the expression of acetylated H3 and H4 histone proteins. In addition, PsA markedly up-regulated the expression of cyclin-dependent kinase inhibitor, p21(WAF1), and down-regulated the expression of pRb, cyclins, and CDKs, which lead to induce cell cycle arrest. Cell cycle analysis indicated that PsA treatment increased the proportion of cells in the G0/G1 and G2/M phases, and decreased the ratio of cells in the S phase.
The PsA treatment resulted in the significant induction of apoptosis, which was associated with p53 independent p21(WAF1) expression. These results suggest that PsA exhibits the antiproliferative effects on endometrial cancer cells through selective induction of genes related to cell cycle arrest and apoptosis.
[show abstract][hide abstract] ABSTRACT: Silibinin is the primary active compound in silymarin. It has been demonstrated to exert anti-carcinogenic effects and hepato-protective effects. However, the effects of silibinin on the maturation and immunostimulatory activities exhibited by dendritic cells (DCs) remain, for the most part, unknown. In this study, we have attempted to determine whether silibinin can influence surface molecule expression, dextran uptake, cytokine production, capacity to induce T-cell differentiation, and the signaling pathways underlying these phenomena in murine bone marrow-derived DCs. Silibinin was shown to significantly suppress the expression of CD80, CD86, MHC class I, and MHC class II in the DCs, and was also associated with impairments of LPS-induced IL-12 expression in the DCs. Silibinin-treated DCs proved highly efficient with regard to Ag capture via mannose receptor-mediated endocytosis. Silibinin also inhibited the LPS-induced activation of MAPKs and the nuclear translocation of the NF-kappaB p65 subunit. Additionally, silibinin-treated DCs evidenced an impaired induction of Th1 response, and a normal cell-mediated immune response. These findings provide new insight into the immunopharmacological functions of silibinin, especially with regard to their impact on the DCs. These findings expand our current understanding of the immunopharmacological functions of silibinin, and may prove useful in the development of therapeutic adjuvants for acute and chronic DC-associated diseases.
Journal of Cellular Physiology 03/2007; 210(2):385-97. · 4.22 Impact Factor
[show abstract][hide abstract] ABSTRACT: Test of human papillomavirus (HPV) is a useful adjunctive tool of Pap smear to screen cervical cancer. We have developed a novel HPV genotyping DNA chip arrayed by multiple oligonucleotide probes of both L1 and E6/E7 gene sequence of 42 types of anogenital HPV.
Consensus PCR products of L1 and E6/E7 gene sequences of HPV are hybridized to arrayed probes on the HPV chip and HPV genotypes are identified by fluorescence scanner. We have comparatively analyzed the value of HPV DNA chip and DNA sequencing in 100 cervical cancer tissues.
Overall, 98 cervical cancer tissues were found to harbor DNA sequences of high-risk type HPVs, of which 88 (89.8%) were detected by PCR-sequencing of L1 alone, 98 (100%) by PCR-sequencing of both L1 and E6/E7, and 98 (100%) by HPV DNA chip, respectively. All of the genotypes of HPV detected on sequencing analysis were also found on DNA chip analysis. HPV DNA chip was superior to direct DNA sequencing in detection of mixed infection.
These results suggest that HPV DNA chip analysis in the present study is highly accurate for detection and genotyping of HPV and may have potential value as a robust, high-throughput screening test of uterine cervix cancer.
[show abstract][hide abstract] ABSTRACT: Our purpose was to establish an evaluation system for oocyte quality based on the incidence of cumulus cells apoptosis and to examine the effect of coculture, using autologous cumulus cells, on the outcome of IVF-ET according to proliferative activities of helper cells and the incidence of cumulus cells apoptosis.
Cumulus cell masses were collected from 91 mature oocytes among 330 oocytes retrieved from a total of 34 IVF-ET cycles with tubal infertility and unexplained infertility. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. On ovum pick up, 2nd day embryos were cocultured with autologous cumulus cells. Prior to coculture, in vitro proliferative activity of cumulus cells was evaluated.
Cumulus cells from patient groups over 40 years old had a significantly increased apoptosis incidence, a lower fertilization rate, and the decreased number of oocytes retrieved compared to the other age groups (P < .05). The incidence of cumulus cells apoptosis was significantly lower when the number of oocytes retrieved was 5 or less (P < .05). Cumulus cells from fertilized oocytes (0.43 +/- 0.07%) and those from patients who became pregnant (0.44 +/- 0.11%) following IVF-ET showed a significantly lower incidence of apoptosis compared to those of unfertilized oocytes (1.80 +/- 0.35%; P < .001) and the nonpregnant group (0.81 +/- 0.10%; P < .05). Embryo quality also had a negative correlation with the incidence of cumulus cells apoptosis. Coculture of fertilized oocytes with cumulus cells with high proliferative activity resulted in improved rates of implantation and pregnancy compared to that with poor active cumulus cells. No significant difference was found between the in vitro proliferative activity of cumulus cells and the incidence of cumulus cells apoptosis (P < .063).
The age of women might influence the incidence of apoptosis in cumulus cells, and the increased incidence of apoptosis is associated with the number of oocytes retrieved, the fertilization rate, and the pregnancy outcome following IVF-ET. These results suggest that the incidence of cumulus cells apoptosis can be used in predicting oocyte quality, outcome of IVF-ET, and age-related decline in fertility.
Journal of Assisted Reproduction and Genetics 10/2001; 18(9):490-8. · 1.82 Impact Factor
[show abstract][hide abstract] ABSTRACT: To elucidate the mechanism for the mode of action of coculture by the use of a coculture system for mouse one-cell embryos with human oviductal epithelial cells.
Prospective, controlled in vitro experimental study.
Academic research laboratory.
Female ICR strain mice aged between 6 and 8 weeks.
Flushed one-cell embryos were cultured in human tubal fluid medium alone (control), in coculture system with human oviductal cells, in five kinds of conditioned media, and in a contactless coculture system using a cell-culture insert.
The percentage of the embryos developed to hatching blastocyst stage and the level of superoxide anion in the supernatant from each culture condition.
The rates of embryo development to the hatching blastocyst stage were significantly higher in the coculture group (43%) than in the control group (none) (P <.05). The embryo development rate in the control group was similar to that of the embryos in the five kinds of conditioned media. The effects of coculture on embryo development disappeared in the contactless coculture group. The level of superoxide anion was significantly reduced in the coculture group compared to the control group.
The present coculture system overcomes the two-cell block in vitro and improves the embryo development. The beneficial effect may be a result of direct cell-to-cell contact between the embryo and helper cells and the removal of deleterious components from medium, rather than a result of embryotrophic factors.
Fertility and Sterility 01/2001; 75(1):193-9. · 4.17 Impact Factor
[show abstract][hide abstract] ABSTRACT: Our objective was to explain a relationship between concentrations of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) in follicular fluid, oocyte quality, and outcomes of in vitro fertilization--embryo transfer (IVF-ET).
The concentrations of TNF-alpha and NO were measured in 115 follicular fluid samples collected from 43 patients undergoing IVF-ET program, due to tubal obstruction, some with endometriosis (8 patients) or hydrosalpinx (5 patients). A correlation of these factors concentrations and the oocyte quality, the oocyte maturity, and infertility-associated diseases was analyzed.
No correlation was found between concentrations of NO and TNF-alpha in follicular fluid. NO concentrations in follicular fluids were significantly higher in patients with endometriosis (P < 0.001) or hydrosalpinx (P < 0.01) compared to the patients with just tubal obstruction. Follicular NO concentration differences according to oocyte maturity and oocyte quality were not found. In contrast, TNF-alpha concentrations in follicular fluids were significantly higher in poor quality oocytes (P < 0.05) but were not associated with infertility-associated diseases, such as hydrosalphinx or endometriosis,and the oocyte maturity. No significant differences in follicular levels of NO and TNF-alpha as well as IVF-ET parameters of pregnant and nonpregnant groups were revealed.
There is no significant correlation between the concentrations of NO and TNF-alpha in follicular fluid. NO levels in follicular fluid are altered in infertility-associated diseases. However, TNF-alpha levels but not NO levels influence oocyte quality. These results suggest that the production of NO and TNF-alpha in follicular fluid may be regulated via different pathways and can be tempered with infertility-associated diseases, thereby influencing oocyte quality locally.
Journal of Assisted Reproduction and Genetics 04/2000; 17(4):222-8. · 1.82 Impact Factor
[show abstract][hide abstract] ABSTRACT: Purpose: Our objective was to explain a relationshipbetween concentrations of tumor necrosis factor- (TNF-)and nitric oxide (NO) in follicular fluid, oocyte quality, andoutcomes of in vitro fertilization=nembryo transfer (IVF-ET).
Methods: The concentrations of TNF- and NO weremeasured in 115 follicular fluid samples collected from 43patients undergoing IVF-ET program, due to tubalobstruction, some with endometriosis (8 patients) or hydrosalpinx(5 patients). A correlation of these factors concentrationsand the oocyte quality, the oocyte maturity, andinfertility-associated diseases was analyzed.
Results: No correlation was found between concentrationsof NO and TNF- in follicular fluid. NO concentrations infollicular fluids were significantly higher in patients withendometriosis (P < 0.001)="" or="" hydrosalpinx="" (p="">< 0.01)compared="" to="" the="" patients="" with="" just="" tubal="" obstruction.="" follicularno="" concentration="" differences="" according="" to="" oocyte="" maturityand="" oocyte="" quality="" were="" not="" found.="" in="" contrast,="">concentrations in follicular fluids were significantly higher inpoor quality oocytes (P < 0.05)="" but="" were="" not="" associatedwith="" infertility-associated="" diseases,="" such="" as="" hydrosalphinxor="" endometriosis,="" and="" the="" oocyte="" maturity.="" no="" significantdifferences="" in="" follicular="" levels="" of="" no="" and=""> as well asIVF-ET parameters of pregnant and nonpregnant groupswere revealed.
Conclusions: There is no significant correlation betweenthe concentrations of NO and TNF- in follicular fluid. NOlevels in follicular fluid are altered in infertility-associateddiseases. However, TNF- levels but not NO levels influenceoocyte quality. These results suggest that the production ofNO and TNF- in follicular fluid may be regulated viadifferent pathways and can be tempered withinfertility-associated diseases, thereby influencing oocyte qualitylocally.
Journal of Assisted Reproduction and Genetics 03/2000; 17(4):222-228. · 1.82 Impact Factor