[Show abstract][Hide abstract] ABSTRACT: We characterised DNA methylation and gene expression of four tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors DR4, DR5, DcR1 and DcR2 in three choriocarcinoma (JAR, JEG-3, BeWo) and two transformed (HTR-8/SVneo and HPT-8) cell lines. DR4 mRNA was detected in JAR, JEG-3, BeWo and HTR-8/SVneo cells, whereas DR5 was present in all detected cells. DcR1 transcripts were expressed only in JAR, JEG-3 and BeWo cells, whereas DcR2 transcripts were detected only in HTR-8/SVneo and HPT-8 cells. Hypermethylated DR4 promoter was observed in JAR, JEG-3, BeWo and HTR-8/SVneo cells, hypermethylated DcR1 promoter in HTR-8/SVneo and HPT-8 cells and hypermethylated DcR2 promoter in JAR, JEG-3 and BeWo cells. Restoration of DR4, DcR1 and DcR2 expression with decreased DNA methylation of these genes was induced by the DNA demethylation agent 5-aza-2'-deoxycytidine (5-aza-CdR) in trophoblast cells, whereas DR5 expression did not exhibit any change. Significant negative correlation between the expression and DNA methylation of these genes was also observed. In all tested cell lines, only HPT-8 demonstrated sensitivity to TRAIL-induced apoptosis. Combined treatment with 5-aza-CdR and TRAIL resulted in apoptosis in JAR, JEG-3, BeWo and HTR-8/SVneo cells but not in HPT-8 cells. The results indicate that DNA methylation is associated with TRAIL receptor expression and might be involved in trophoblast apoptosis.
Preview · Article · May 2015 · Reproduction Fertility and Development
[Show abstract][Hide abstract] ABSTRACT: The racial/ethnic disparities in DNA methylation patterns indicate that molecular markers may play a role in determining the individual susceptibility to diseases in different ethnic groups. Racial disparities in DNA methylation patterns have been identified in prostate cancer, breast cancer and colorectal cancer and are related to racial differences in cancer prognosis and survival.
No preview · Article · Jul 2014 · Biochimica et Biophysica Acta (BBA) - Reviews on Cancer
[Show abstract][Hide abstract] ABSTRACT: Trisomy 21 is a chromosomal condition caused by the presence of all or part of an extra 21st chromosome. There has been limited research into the DNA methylation status of CpG islands (CGIs) in trisomy 21, therefore, exploring the DNA methylation status of CGIs in 21q is essential for the development of a series of potential epigenetic biomarkers for prenatal screening of trisomy 21. First, DNA sequences of CGIs in 21q from the USCS database were obtained and 149 sequences and 148 pairs of primers in the BGI YH database were aligned. All 300 cases were analyzed by a heavy methyl‑polymerase chain reaction (HM‑PCR) assay and a comparison of the DNA methylation status of CGIs was made between trisomy 21 and the control. The HM‑PCR assay results did not show a difference in the DNA methylation status between individuals with trisomy 21 and the control. In total, there were 11 CGIs that showed various DNA methylation statuses between Japanese and Chinese patients. Subsequently, bisulfite genomic sequencing found variations in the methylation status of CpG dinucleotides in CGIs (nos. 14, 75, 109, 134 and 146) between trisomy 21 and the control. The different DNA methylation status of CpG dinucleotides in CGIs may be a potential epigenetic marker for diagnosing trisomy 21. No difference was identified in the DNA methylation status of 21q CGIs among Chinese individuals with trisomy 21 and the control. The homogeneity of the DNA methylation status of 21q CGIs in Chinese patients indicates that DNA methylation is likely to be an epigenetic marker distinguishing ethnicities.
No preview · Article · Feb 2014 · Molecular Medicine Reports
[Show abstract][Hide abstract] ABSTRACT: To study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation.
The expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR.
IDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γ increased its expression.
The expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.
No preview · Article · Nov 2013 · Nan fang yi ke da xue xue bao = Journal of Southern Medical University
[Show abstract][Hide abstract] ABSTRACT: Benzo(a)pyrene (B[a]P) is an environmental carcinogen that induces tumors in many animal species, but the neurotoxic effects of B[a]P have not been well studied. In the present study, we investigated the effects of subacute exposure to B[a]P in Sprague-Dawley (SD) rats. Male rats received daily injections of either B[a]P (0, 1, 2.5, or 6.25mg/kg, i.p.) or vehicle for 45 days. Exposure to B[a]P affected the behavior of rats in the Morris water maze test. Gene microarray and real-time PCR analyses revealed that exposure to B[a]P affected signal transduction in the rat hippocampus. Protein microarray analysis revealed that altered protein expression played a role in cell death in the functional annotation cluster analysis. Finally, major vault protein was found to display low cDNA and protein expression levels. The present study explored some of the possible mechanisms underlying B[a]P neurotoxicity and provided evidence that B[a]P plays a neurotoxic role in rats.
[Show abstract][Hide abstract] ABSTRACT: Successful mouse embryo implantation requires a receptive uterus and an activated blastocyst. A large number of genes, cytokines, and other factors are involved in the process. MicroRNAs (miRNAs) regulate the expression of many genes, and previous studies have investigated the relationship between miRNA expression and embryo implantation. In this study, we show that mmu-microRNA-200a (mmu-miR-200a) is expressed in a spatiatemporal manner during implantation in mouse uterus and found that phosphatase and tensin homolog (PTEN), SON, and programmed cell death 4 (Pdcd4) are the target genes of mmu-miR-200a by bioinformatics analysis. In vitro gain and loss of function experiments confirm that PTEN, a critical gene for cell proliferation and apoptosis, is the target gene of mmu-miR-200a. Our experiments also show that injection of the uterine horn with mmu-miR-200a lentivirus leads to a decreased implantation rate. Collectively, our results suggest that mmu-miR-200a affects embryo implantation by regulating PTEN protein expression. Thus, clarifying the physiological functions of uterine miRNAs will help to elucidate the embryo implantation process and may even contribute to curing infertility and inventing new contraceptives.
No preview · Article · May 2013 · Reproductive sciences (Thousand Oaks, Calif.)
[Show abstract][Hide abstract] ABSTRACT: The 'fetal origin hypothesis' suggests that metabolic diseases are directly related to poor nutritional status in early life. Thus, a high birth weight (HBW) may pose a lower risk than normal birth weight. Overweight and overnutrition are among the most widely recognized risk factors of metabolic diseases. To explore the possible effects of HBW on blood pressure and hypertension, a systematic review was performed. The PubMed and Embase databases were searched for relevant studies. The outcomes included systolic blood pressure (SBP), diastolic blood pressure (DBP) and hypertension. We included all of the studies that assessed the differences in outcomes for children aged >1 year between those born with normal birth weight (birth weight between 2500 and 4000 g or between the 10th and 90th percentiles for their gestational age) and those born with HBW (birth weight4000 g or 90th percentile for their gestational age). The outcomes were analyzed descriptively and by conducting a meta-analysis. Thirty-one studies satisfied the inclusion criteria. The mean difference in blood pressure and the relative risk of hypertension between individuals with HBW and individuals with normal birth weight was inversely associated with age. SBP and DBP, as well as the prevalence of hypertension, were higher in younger children with HBW but lower in older adults with HBW compared with individuals with normal birth weight. The findings suggested that an individual with HBW is prone to hypertension and higher blood pressure during childhood. However, a 'catch-down' effect in the elevation of blood pressure is observed in subjects with HBW as they grow older. Thus, older individuals with HBW are less susceptible to hypertension than those with normal birth weight.Hypertension Research advance online publication, 18 April 2013; doi:10.1038/hr.2013.33.
Full-text · Article · Apr 2013 · Hypertension Research
[Show abstract][Hide abstract] ABSTRACT: Full development of a receptive uterus is necessary for embryo implantation; however, many genes that are required for the endometrial modifications that occur during this process remain unidentified. To identify novel genes that control endometrial modifications during this period, we investigated the differential gene expression profile in the endometrium of mice on days 2 (D2) (pre-implantation) and 4 (D4) of pregnancy (i.e., the implantation window) using 17-bp long serial analysis of gene expression (LongSAGE). One hundred fifty-six tags were annotated as unique transcripts. Of these, 101 tags were significantly upregulated, and 55 tags were downregulated in the D4 library relative to the D2 library. These differentially expressed genes should therefore be of increased importance in the establishment of uterine receptivity. The differential expressions of certain of the identified genes, namely, Hspa8, Tctp, Sparc, Ifitm1, Ik, serbp1 and Dnmt1, were validated by semi-quantitative RT-PCR and/or immunohistochemistry. Functional grouping analysis classified 86 of the mapped tags into 17 categories, which are closely associated with morphological modifications of the endometrium during pregnancy. Ingenuity pathways analysis revealed that the identified differentially expressed genes fell into six primary networks, which themselves contain numerous factors that are related to key modulators of signaling pathways that are vital for endometrial modifications. These findings will aid in the further understanding of the molecular events that underlie the implantation physiology in mice.
No preview · Article · Oct 2012 · Molecular Biology Reports
[Show abstract][Hide abstract] ABSTRACT: The DNA demethylating agent 5-aza-2'-deoxycytidine (5-aza-CdR) incorporates into DNA and decreases DNA methylation, sparking interest in its use as a potential therapeutic agent. We aimed to determine the effects of maternal 5-aza-CdR treatment on embryo implantation in the mouse and to evaluate whether these effects are associated with decreased levels of DNA methyltransferases (Dnmts) and three genes (estrogen receptor α [Esr1], progesterone receptor [Pgr], and homeobox A10 [Hoxa10]) that are vital for control of endometrial changes during implantation.
Mice treated with 5-aza-CdR had a dose-dependent decrease in number of implantation sites, with defected endometrial decidualization and stromal cell proliferation. Western blot analysis on pseudo-pregnant day 3 (PD3) showed that 0.1 mg/kg 5-aza-CdR significantly repressed Dnmt3a protein level, and 0.5 mg/kg 5-aza-CdR significantly repressed Dnmt1, Dnmt3a, and Dnmt3b protein levels in the endometrium. On PD5, mice showed significantly decreased Dnmt3a protein level with 0.1 mg/kg 5-aza-CdR, and significantly decreased Dnmt1 and Dnmt3a with 0.5 mg/kg 5-aza-CdR. Immunohistochemical staining showed that 5-aza-CdR repressed DNMT expression in a cell type-specific fashion within the uterus, including decreased expression of Dnmt1 in luminal and/or glandular epithelium and of Dnmt3a and Dnmt3b in stroma. Furthermore, the 5' flanking regions of the Esr1, Pgr, and Hoxa10 were hypomethylated on PD5. Interestingly, the higher (0.5 mg/kg) dose of 5-aza-CdR decreased protein expression of Esr1, Pgr, and Hoxa10 in the endometrium on PD5 in both methylation-dependent and methylation-independent manners.
The effects of 5-aza-CdR on embryo implantation in mice were associated with altered expression of endometrial Dnmts and genes controlling endometrial changes, suggesting that altered gene methylation, and not cytotoxicity alone, contributes to implantation defects induced by 5-aza-CdR.
[Show abstract][Hide abstract] ABSTRACT: Tubeimoside-1 (TBMS1) extracted from Bolbostemma paniculatum (Maxim), is a traditional Chinese herb with anticancer potential. It induces apoptosis in a number of human carcinoma cell lines, but the mechanism has remained unclear. In the present study, we investigated the pro-apoptotic activity of TBMS1 against SKOV-3 cell lines and the underlying mechanisms. Treatment with TBMS1 resulted in dose- and time-dependent inhibition of proliferation, led to arrest in phase G2/M of the cell cycle and increased the levels of intracellular Ca²⁺. Furthermore, TBMS1 up-regulated the levels of the glucose-regulated protein 78/immunoglobuin heavy chain binding protein (GRP78/Bip), C/EBP homologous protein (CHOP), Bax, and cleaved caspase-3 and down-regulated the levels of Bcl-2. It was shown to be linked to activation of the extracellular signal-regulated kinase (ERK) 1 and 2 signal transduction pathway. A decrease in Bcl-2/Bax ratio with increased expression of caspase-3, and intracellular Ca²⁺ provide compelling evidence that TBMS1-induced apoptosis is mediated by the mitochondrial pathway. The results of the present study suggest that TBMS1 has immense potential in cancer prevention and therapy based on its antiproliferative and apoptosis inducing effects.
Preview · Article · Feb 2012 · International Journal of Oncology
[Show abstract][Hide abstract] ABSTRACT: To assess the relationship between folate biomarkers levels and IGF2 imprinting status among second trimester pregnant Chinese women.
Three hundred women in their second trimester were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for analysis of IGF2 imprinting status. Statistical differences of folate biomarkers levels were calculated in the groups with different imprinting status.
Of the 300 women analyzed, 133 (44.33%) cases were homozygous allele with A, 3 (1.00%) cases were homozygous allele with G, while 164 (54.67%) cases were heterozygous with A and G, qualifying them for analysis of loss of imprinting (LOI). Among the 164 cases undergoing LOI analysis, 44 (26.83%) were IGF2 LOI cases, while 120 (73.17%) were IGF2 retention of imprinting (ROI) ones. The mean level of serum folate, vitamin B12 and tHcy was 28.46 +/- 10.74 ng/mL, 380.20 +/- 206.13 pg/mL, 14.24 +/- 6.34 micromol/L among women with IGF2 ROI, and 30.89 +/- 9.97 ng/mL, 394.28 +/- 195.92 pg/ mL, and 13.12 +/- 6.23 micromol/L among women with IGF2 LOI, respectively.
No significant difference of folate biomarkers levels was observed between IGF2 ROI and IGF2 LOI groups.
No preview · Article · Dec 2011 · The Journal of reproductive medicine
[Show abstract][Hide abstract] ABSTRACT: To explore the effects of the death receptor (mouse killer, MK) of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on decidualization of mouse uterine stromal cells, both of the over-expression and siRNA of MK gene recombinant adenovirus were applied to infect the primary culture of mouse uterine stromal cells followed by decidualization induction with estrogen, progesterone plus cAMP. Seventy-two hours later, immunocytochemical assay and flow cytometry were used to detect the prolactin protein expression and apoptosis of decidualized cells, which were transfected with MK recombinant adenovirus (overexpression and RNAi). Moreover, each MK recombinant adenovirus was injected into the uterine horns of mice on the early morning of d4 of pregnancy. The number of implanting embryos was counted on d8 of pregnancy. The results showed that the levels of prolactin protein decreased significantly, but the apoptosis rate increased in the decidualized cells transfected with MK over-expression adenovirus. In the MK siRNA group, the prolatin levels were increased while the apoptosis rate was downregulated markedly. However, uterine injection of either the over-expression or the siRNA adenovirus led to dramatic decline the numbers of the implanting embryos. These results suggested that MK was involved in modulating the decidulization of mouse uterine stromal cells possibly through balancing proliferation and apoptosis of the decidualized cells.
No preview · Article · Aug 2011 · Progress in Biochemistry and Biophysics
[Show abstract][Hide abstract] ABSTRACT: Mitochondria play important roles in the intrinsic pathways that trigger apoptosis. Anticancer chemotherapies eliminate cancer cells mainly through the induction of apoptosis. In the present study, we investigated the mechanism of the cytotoxic effects of tubeimoside I (TBMS1) on the human choriocarcinoma cell line (JEG-3). Choriocarcinoma is one of the most common malignant tumors in the reproductive system. TBMS1, a triterpenoid saponin, isolated from the tubers of Bolbostemma paniculatum (Maxim) Franquet (Cucurbitaceae), showed potent antitumor effects. However, the potential roles of TBMS1 in the treatment of choriocarcinoma remain unknown. In the present study, we examined the effects of TBMS1 on JEG-3 cells. TBMS1 displayed strong growth inhibitory effects on JEG-3 cell growth. In addition, TBMS1 treatment with TBMS1 led to marked cell apoptosis, significant cell cycle arrest at G2 phase and decrease in mitochondrial transmembrane potential (ΔΨm). Cytochrome c was released from the mitochondria and caspase-3 expression was enhanced. Furthermore, TBMS1 induced the up-regulation of Bcl-2 associated X protein (Bax) expression, down-regulation of Bcl-2 expression, inhibition of nuclear factor-κ-B (NF-κB) function and impacted the phosphorylation of p38 mitogen-activated protein kinase (p38/MAPK), extracellular signal-regulated kinases (ERK)1/2 and protein kinase B (Akt). Taken together, our findings suggest that TBMS1 is an efficient apoptosis-inducing agent for choriocarcinoma cells, which exerts its effects, at least partially, by the induction of mitochondrial dysfunction and regulation of the p38/MAPK, ERK1/2 and PI3K/Akt signaling pathways.
No preview · Article · Jun 2011 · International Journal of Molecular Medicine
[Show abstract][Hide abstract] ABSTRACT: Cisplatin (CDDP) is a major chemotherapeutic drug used in the treatment of human ovarian cancer. Tubeimoside I (TBMS1) has also shown potent antitumor and antitumor-promoting effects, and may offer a promising new approach in the effective treatment of CDDP-resistant human ovarian cancers. This study aimed to investigate the effect of TBMS1 in sensitizing CDDP in CDDP-resistant human ovarian cancer cells (A2780/DDP). A variety of methods were employed to measure cell apoptosis, p38, ERK1/2 and glutathione S-transferase (GST)-π expressions. It was found that TBMS1 combined with CDDP promoted cell apoptosis, decreased proliferation activity and increased cytosolic Ca2+ levels. Bcl-2 protein expression was down-regulated but Bax was up-regulated. Moreover, GST-π mRNA and protein expression were decreased. TBMS1 reduced the resistance of the cells to CDDP-induced cytotoxicity. Both the p38 inhibitor (SB203580) and the ERK1/2 inhibitor (PD98059) effectively blocked this effect. These results suggest that TBMS1 can effectively sensitize CDDP in CDDP-resistant human ovarian cancer cells through the down-regulation of the ERK1/2 and the up-regulation of the p38 signaling pathways.
Preview · Article · Jun 2011 · Molecular Medicine Reports
[Show abstract][Hide abstract] ABSTRACT: Interleukin-8 (IL-8) expression by melanoma cells may influence their metastatic capabilities. Tetramethylpyrazine (TMP) from Ligusticum wallichil Franch. possesses anti-inflammatory and antitumor activities. It has recently been suggested that autocrine IL-8 may play a role in tumor cell survival, invasion and migration. The role of TMP in association with IL-8 in the tumor cell migratory process remains unclear. The purpose of the present study was to determine whether TMP influences the migratory ability of a human ovarian carcinoma cell line (SKOV3) via regulation of IL-8 expression in vitro. Cell counts showed that treatment of SKOV3 with TMP (25-100 µg/ml) for 24 h did not decrease cell numbers, while an effect of TMP on the down-regulation of the expression of IL-8 was observed. In addition, migration of SKOV3 cells was suppressed after treatment with TMP (25-100 µg/ml) for 24 h. Therefore, expression of IL-8 by SKOV3 cells correlates with their metastatic potential. Western blot analysis revealed that ERK1/2 and p38 phosphorylation was blocked by TMP. Furthermore, IL-8 mRNA expression was inhibited significantly after co-incubation with PD98059 (ERK inhibitor) and SB203580 (p38 inhibitor), respectively. Notably, these changes were the results of activator protein-1 (AP-1) activity suppression rather than that of NF-κB. Our data suggest that TMP may inhibit tumor cell invasion and migration, at least in part, through its down-regulation of IL-8 expression. Our results provide evidence that anti-inflammation plays an important role in integrative cancer therapies.
No preview · Article · May 2011 · Oncology Reports
[Show abstract][Hide abstract] ABSTRACT: As a highly conserved nuclear protein, death domain-associated protein (Daxx) plays an important role in transcriptional control, carcinogenesis, and resistance to virus infection and so on. In order to further investigate the mechanism of Daxx, the yeast two-hybrid technique was used to screen the intra-cellular proteins interacting with Daxx. And 13 positive colonies and three proteins interacting with Daxx were obtained. One of the candidate proteins was identified as ferritin, heavy polypeptide 1(FTH1). The interaction between Daxx and FTH1 was further supported by GST pull-down and co-immunoprecipitation respectively. Then Daxx was determined to induce apoptosis and FTH1 can inhibit Daxx-mediated apoptosis. Besides, it is found that Daxx mediated apoptosis through the Fas-Daxx-ASK1-JNK1 signaling pathway, while FTH1 can inhibit the activation of JNK signaling pathway. We present evidence to demonstrate the FTH1 and Daxx are able to participate in apoptosis pathway through JNK signal molecule and FTH1 can inhibit this pathway.
No preview · Article · May 2011 · Molecular Biology Reports
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to investigate the inhibitory effect of geniposide on lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) and interleukin-8 (IL-8) production in human umbilical vein endothelial cells (HUVECs).
Primary HUVECs were used. The mRNA/protein levels of IL-6 and IL-8 was determined by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). LPS-induced HUVEC migration and adhesion of monocytes to HUVECs were studied by monolayer wound healing experiments and monocytic cell adhesion assay, respectively. Expression of nuclear factor kappaB (NF-kappaB), inhibitory factor kappaB-alpha (IkappaB-alpha), p38 mitogen-activated protein kinase (MAPK) and ERK1/2 were determined by Western blot analysis.
Geniposide effectively inhibited LPS-induced expression of IL-6 and IL-8 in HUVECs at the transcription and translation levels. Additionally, geniposide suppressed LPS-induced HUVEC migration and U937 monocyte adhesion to HUVECs. Signal transduction studies indicate that geniposide blocked the activation of NF-kappaB, degradation of IkappaB-alpha, and phosphorylation of p38 MAPK and ERK1/2 in HUVECs challenged by LPS.
The results show that geniposide can inhibit LPS-induced IL-6 and IL-8 production in HUVECs by blocking p38 MAPK and ERK1/2 signaling pathways.
No preview · Article · Jun 2010 · Agents and Actions
[Show abstract][Hide abstract] ABSTRACT: This study aimed to investigate the protective effect of chitosan oligosaccharides (COS) on human umbilical vein endothelial cell (HUVEC) apoptosis induced by hydrogen peroxide (H2O2). We found that COS not only reversed the decrease of cell viability and proliferation activity, but ameliorated nuclear chromatin damage in H2O2-induced HUVECs. Additionally, COS contributed to the decrease of cytosolic Ca2+ level, increase of mitochondrial membrane potential, up-regulation of Bcl-2 mRNA, down-regulation of Bax mRNA, reduction of cleaved caspase-3 protein, inhibition of phosphorylated p38 mitogen-activated protein kinase (MAPK) and induction of phosphorylated Akt in HUVECs. Further, H2O2-induced caspase-3 activation could be suppressed by p38 MAPK inhibitor (SB203580) and up-regulated by phosphatidylinositol-3-kinase (PI3K) inhibitor (LY294002), indicating the involvement of p38 MAPK and PI3K/Akt signaling pathways. In conclusion, the results show that COS may effectively inhibit the H2O2-induced HUVEC apoptosis.
Full-text · Article · May 2010 · Carbohydrate Polymers