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Vadivel Ganapathy - One of the best experts on this subject based on the ideXlab platform.

  • human placental na dependent multivitamin transporter cloning functional expression gene structure and chromosomal localization
    Journal of Biological Chemistry, 1999
    Co-Authors: Haiping Wang, You Jun Fei, Teresa L Yangfeng, Frederick H Leibach, Vadivel Ganapathy, Wei Huang, Hong Xia, Lawrence D Devoe, Puttur D. Prasad
    Abstract:

    We have cloned the human Na+-dependent multivitamin transporter (SMVT), which transports the water-soluble vitamins pantothenate, biotin, and lipoate, from a placental Choriocarcinoma Cell Line (JAR). The cDNA codes for a protein of 635 amino acids with 12 transmembrane domains and 4 putative sites for N-linked glycosylation. The human SMVT exhibits a high degree of homology (84% identity and 89% similarity) to the rat counterpart. When expressed in HRPE Cells, the cDNA-induced transport process is obligatorily dependent on Na+ and accepts pantothenate, biotin, and lipoate as substrates. The relationship between the cDNA-specific uptake rate of pantothenate or biotin and Na+ concentration is sigmoidal with a Na+:vitamin stoichiometry of 2:1. The human SMVT, when expressed in Xenopus laevis oocytes, induces inward currents in the presence of pantothenate, biotin, and lipoate in a Na+-, concentration-, and potential-dependent manner. We also report here on the structural organization and chromosomal localization of the human SMVT gene. TheSMVT gene is ∼14 kilobase pairs in length and consists of 17 exons. The SMVT gene is located on chromosome 2p23 as evidenced by somatic Cell hybrid analysis and fluorescence in situ hybridization.

  • transport mechanisms for vitamin c in the jar human placental Choriocarcinoma Cell Line
    Biochimica et Biophysica Acta, 1998
    Co-Authors: Puttur D. Prasad, Frederick H Leibach, Haiping Wang, Wei Huang, Vadivel Ganapathy
    Abstract:

    We investigated the transport pathways available for the uptake of vitamin C in the human placental Choriocarcinoma Cell Line, JAR. These Cells were found to possess the capacity to accumulate the vitamin when presented either in the oxidized form (dehydroascorbic acid) or in the reduced form (ascorbate). Dithiothreitol and 5,5′-dithiobis(2-nitrobenzoic acid) were used to maintain vitamin C as ascorbate and dehydroascorbic acid, respectively. The uptake of these two forms of vitamin C in JAR Cells was found to occur by different mechanisms. The uptake of the dehydroascorbic acid was Na+-independent and was mediated by facilitative glucose transporters as evidenced from the inhibition of the uptake process by glucose. On the other hand, the uptake of ascorbate was Na+-dependent and was not sensitive to inhibition by glucose. Substitution of Na+ with other monovalent cations abolished the uptake of ascorbate completely. The uptake process was, however, not influenced by anions. Kinetic analysis indicated the presence of a single saturable transport system for ascorbate with a Michaelis-Menten constant of 22 ± 1μM. The dependence of the uptake rate of ascorbate on Na+ concentration exhibited sigmoidal kinetics, suggesting interaction of more than one Na+ ion with the transporter. The Hill coefficient for the Na+ interaction was 2, indicating that the Na+-dependent ascorbate transport is electrogenic. The Na+-dependent stimulation of ascorbate uptake was primarily due to an increase in the affinity of the transporter for ascorbate in the presence of Na+. It is concluded that the JAR placental trophoblast Cell Line expresses two different transport systems for vitamin C: one for the reduced form of the vitamin ascorbate; and the other for the oxidized form of the vitamin dehydroascorbic acid.

  • tyrosine phosphorylation and epidermal growth factor dependent regulation of the sodium coupled amino acid transporter b0 in the human placental Choriocarcinoma Cell Line jar
    Biochimica et Biophysica Acta, 1997
    Co-Authors: Viviana Torreszamorano, Ramesh Kekuda, Frederick H Leibach, Vadivel Ganapathy
    Abstract:

    We have recently cloned an amino acid transporter from the human placental Choriocarcinoma Cell Line JAR which, when functionally expressed in HeLa Cells, induces an amino acid transport activity with characteristics known to be associated with the amino acid transport system B(0) (R. Kekuda, P.D. Prasad, Y.J. Fei, V. Torres-Zamorano, S. Sinha, T.L. Yang-Feng, F.H. Leibach, and V. Ganapathy, J. Biol. Chem. 271, 18657-18661, 1996). The presence of the amino acid transport system B(0) (ATB(0)) has however not been previously described in these Cells by functional studies. In the present investigation, we have obtained evidence for the existence of ATB(0) in JAR Cells and deLineated the functional characteristics of the transporter. The identifying characteristics include Na(+)-dependence and preference for neutral amino acids. In addition, we have used the JAR Cells as a model system to investigate the regulatory aspects of ATB(0). Treatment of the Cells with the neuroprotective agent aurintricarboxylic acid (ATA) for 16 h leads to a significant increase in ATB(0) activity. This increase is associated with enhanced maximal velocity of the transporter and with increased steady state levels of the transporter mRNA. The effect of ATA is blocked by the tyrosine kinase inhibitor genistein. ATA treatment results in increased tyrosine phosphorylation of two major proteins, 180 kDa and 140 kDa in size. The 180 kDa protein is likely to be the epidermal growth factor (EGF) receptor because exposure of the Cells to EGF also leads to enhanced tyrosine phosphorylation of a protein of similar molecular size. Furthermore, the effects of ATA on ATB(0) activity and on ATB(0) mRNA levels can be reproduced by EGF. Treatment of the Cells with EGF for 24 h results in a significant increase in ATB(0) activity and this effect is associated with an increase in the maximal velocity of the transporter and with an increase in the steady state levels of the transporter mRNA. These data suggest that ATA influences ATB(0) activity in JAR Cells most likely by activating the EGF receptor through tyrosine phosphorylation. It is concluded that the human placental Choriocarcinoma Cells functionally express the amino acid transport system B(0) and that the expression of the system in these Cells is stimulated by EGF.

  • cloning of the sodium dependent broad scope neutral amino acid transporter bo from a human placental Choriocarcinoma Cell Line
    Journal of Biological Chemistry, 1996
    Co-Authors: Ramesh Kekuda, Puttur D. Prasad, You Jun Fei, Viviana Torreszamorano, Srish Sinha, Teresa L Yangfeng, Frederick H Leibach, Vadivel Ganapathy
    Abstract:

    We have isolated a cDNA from a human placental Choriocarcinoma Cell cDNA library which, when expressed in HeLa Cells, induces a Na+-dependent amino acid transport system with preference for zwitterionic amino acids. Anionic amino acids, cationic amino acids, imino acids, and N-methylated amino acids are excluded by this system. These characteristics are identical to those described for the amino acid transporter Bo. When expressed in Xenopus laevis oocytes that do not have detectable endogenous activity of the amino acid transporter Bo, the cloned transporter increases alanine transport in the oocytes severalfold and induces alanine-evoked inward currents in the presence of Na+. The cDNA codes for a polypeptide containing 541 amino acids with 10 putative transmembrane domains. Amino acid sequence homology predicts this transporter (hATBo) to be a member of a superfamily consisting of the glutamate transporters, the neutral amino acid transport system ASCT, and the insulin-activable neutral/anionic amino acid transporter. Chromosomal assignment studies with somatic Cell hybrid analysis and fluorescent in situ hybridization have located the ATBo gene to human chromosome 19q13.3.

  • modulation of the activity of amino acid transport system l by phorbol esters and calmodulin antagonists in a human placental Choriocarcinoma Cell Line
    Biochimica et Biophysica Acta, 1992
    Co-Authors: Sammanda Ramamoorthy, Frederick H Leibach, Virendra B. Mahesh, Vadivel Ganapathy
    Abstract:

    We investigated the regulation of the activity of amino acid transport system L in the JAR human placental Choriocarcinoma Cell Line by agents which are known to modulate the activities of three different classes of protein kinases, A-kinase, C-kinase and CaM-kinase. The system L activity was measured by determining the uptake of leucine in these Cells, grown as confluent monolayers. Leucine uptake in these Cells was predominantly Na(+)-independent, was stimulated by lowering the extraCellular pH and was inhibited by hydrophobic neutral amino acids. These characteristics demonstrate that uptake of leucine in this Cell Line occurs primarily via system L. Treatment of the Cells with cholera toxin and forskolin, agents which are known to elevate intraCellular cAMP levels, did not have any effect on the activity of system L. 4 beta-Phorbol 12-myristate 13-acetate, an activator of C-kinase, but not the inactive analogue 4 alpha-phorbol 12,13-didecanoate, caused a significant stimulation of system L. The involvement of C-kinase in the phorbol ester-induced stimulation was supported by the finding that staurosporine, an inhibitor of C-kinase, effectively blocked the stimulation. Calmodulin antagonists, calmidazolium, W-7 and CGS 9343 B stimulated system L activity markedly. The potency of these antagonists was in the following order: calmidazolium greater than CGS 9343 B greater than W-7. This stimulatory effect was specific for system L because systems A and ASC were not stimulated by these agents. The stimulation caused by these agents was primarily due to an increase in the maximal velocity, the apparent Km of the system being only minimally affected. It is concluded that the activity of amino acid transport system L in the JAR human placental Choriocarcinoma Cell Line is stimulated by C-kinase, inhibited by CaM-kinase and unaffected by A-kinase.

Frederick H Leibach - One of the best experts on this subject based on the ideXlab platform.

  • human placental na dependent multivitamin transporter cloning functional expression gene structure and chromosomal localization
    Journal of Biological Chemistry, 1999
    Co-Authors: Haiping Wang, You Jun Fei, Teresa L Yangfeng, Frederick H Leibach, Vadivel Ganapathy, Wei Huang, Hong Xia, Lawrence D Devoe, Puttur D. Prasad
    Abstract:

    We have cloned the human Na+-dependent multivitamin transporter (SMVT), which transports the water-soluble vitamins pantothenate, biotin, and lipoate, from a placental Choriocarcinoma Cell Line (JAR). The cDNA codes for a protein of 635 amino acids with 12 transmembrane domains and 4 putative sites for N-linked glycosylation. The human SMVT exhibits a high degree of homology (84% identity and 89% similarity) to the rat counterpart. When expressed in HRPE Cells, the cDNA-induced transport process is obligatorily dependent on Na+ and accepts pantothenate, biotin, and lipoate as substrates. The relationship between the cDNA-specific uptake rate of pantothenate or biotin and Na+ concentration is sigmoidal with a Na+:vitamin stoichiometry of 2:1. The human SMVT, when expressed in Xenopus laevis oocytes, induces inward currents in the presence of pantothenate, biotin, and lipoate in a Na+-, concentration-, and potential-dependent manner. We also report here on the structural organization and chromosomal localization of the human SMVT gene. TheSMVT gene is ∼14 kilobase pairs in length and consists of 17 exons. The SMVT gene is located on chromosome 2p23 as evidenced by somatic Cell hybrid analysis and fluorescence in situ hybridization.

  • transport mechanisms for vitamin c in the jar human placental Choriocarcinoma Cell Line
    Biochimica et Biophysica Acta, 1998
    Co-Authors: Puttur D. Prasad, Frederick H Leibach, Haiping Wang, Wei Huang, Vadivel Ganapathy
    Abstract:

    We investigated the transport pathways available for the uptake of vitamin C in the human placental Choriocarcinoma Cell Line, JAR. These Cells were found to possess the capacity to accumulate the vitamin when presented either in the oxidized form (dehydroascorbic acid) or in the reduced form (ascorbate). Dithiothreitol and 5,5′-dithiobis(2-nitrobenzoic acid) were used to maintain vitamin C as ascorbate and dehydroascorbic acid, respectively. The uptake of these two forms of vitamin C in JAR Cells was found to occur by different mechanisms. The uptake of the dehydroascorbic acid was Na+-independent and was mediated by facilitative glucose transporters as evidenced from the inhibition of the uptake process by glucose. On the other hand, the uptake of ascorbate was Na+-dependent and was not sensitive to inhibition by glucose. Substitution of Na+ with other monovalent cations abolished the uptake of ascorbate completely. The uptake process was, however, not influenced by anions. Kinetic analysis indicated the presence of a single saturable transport system for ascorbate with a Michaelis-Menten constant of 22 ± 1μM. The dependence of the uptake rate of ascorbate on Na+ concentration exhibited sigmoidal kinetics, suggesting interaction of more than one Na+ ion with the transporter. The Hill coefficient for the Na+ interaction was 2, indicating that the Na+-dependent ascorbate transport is electrogenic. The Na+-dependent stimulation of ascorbate uptake was primarily due to an increase in the affinity of the transporter for ascorbate in the presence of Na+. It is concluded that the JAR placental trophoblast Cell Line expresses two different transport systems for vitamin C: one for the reduced form of the vitamin ascorbate; and the other for the oxidized form of the vitamin dehydroascorbic acid.

  • tyrosine phosphorylation and epidermal growth factor dependent regulation of the sodium coupled amino acid transporter b0 in the human placental Choriocarcinoma Cell Line jar
    Biochimica et Biophysica Acta, 1997
    Co-Authors: Viviana Torreszamorano, Ramesh Kekuda, Frederick H Leibach, Vadivel Ganapathy
    Abstract:

    We have recently cloned an amino acid transporter from the human placental Choriocarcinoma Cell Line JAR which, when functionally expressed in HeLa Cells, induces an amino acid transport activity with characteristics known to be associated with the amino acid transport system B(0) (R. Kekuda, P.D. Prasad, Y.J. Fei, V. Torres-Zamorano, S. Sinha, T.L. Yang-Feng, F.H. Leibach, and V. Ganapathy, J. Biol. Chem. 271, 18657-18661, 1996). The presence of the amino acid transport system B(0) (ATB(0)) has however not been previously described in these Cells by functional studies. In the present investigation, we have obtained evidence for the existence of ATB(0) in JAR Cells and deLineated the functional characteristics of the transporter. The identifying characteristics include Na(+)-dependence and preference for neutral amino acids. In addition, we have used the JAR Cells as a model system to investigate the regulatory aspects of ATB(0). Treatment of the Cells with the neuroprotective agent aurintricarboxylic acid (ATA) for 16 h leads to a significant increase in ATB(0) activity. This increase is associated with enhanced maximal velocity of the transporter and with increased steady state levels of the transporter mRNA. The effect of ATA is blocked by the tyrosine kinase inhibitor genistein. ATA treatment results in increased tyrosine phosphorylation of two major proteins, 180 kDa and 140 kDa in size. The 180 kDa protein is likely to be the epidermal growth factor (EGF) receptor because exposure of the Cells to EGF also leads to enhanced tyrosine phosphorylation of a protein of similar molecular size. Furthermore, the effects of ATA on ATB(0) activity and on ATB(0) mRNA levels can be reproduced by EGF. Treatment of the Cells with EGF for 24 h results in a significant increase in ATB(0) activity and this effect is associated with an increase in the maximal velocity of the transporter and with an increase in the steady state levels of the transporter mRNA. These data suggest that ATA influences ATB(0) activity in JAR Cells most likely by activating the EGF receptor through tyrosine phosphorylation. It is concluded that the human placental Choriocarcinoma Cells functionally express the amino acid transport system B(0) and that the expression of the system in these Cells is stimulated by EGF.

  • cloning of the sodium dependent broad scope neutral amino acid transporter bo from a human placental Choriocarcinoma Cell Line
    Journal of Biological Chemistry, 1996
    Co-Authors: Ramesh Kekuda, Puttur D. Prasad, You Jun Fei, Viviana Torreszamorano, Srish Sinha, Teresa L Yangfeng, Frederick H Leibach, Vadivel Ganapathy
    Abstract:

    We have isolated a cDNA from a human placental Choriocarcinoma Cell cDNA library which, when expressed in HeLa Cells, induces a Na+-dependent amino acid transport system with preference for zwitterionic amino acids. Anionic amino acids, cationic amino acids, imino acids, and N-methylated amino acids are excluded by this system. These characteristics are identical to those described for the amino acid transporter Bo. When expressed in Xenopus laevis oocytes that do not have detectable endogenous activity of the amino acid transporter Bo, the cloned transporter increases alanine transport in the oocytes severalfold and induces alanine-evoked inward currents in the presence of Na+. The cDNA codes for a polypeptide containing 541 amino acids with 10 putative transmembrane domains. Amino acid sequence homology predicts this transporter (hATBo) to be a member of a superfamily consisting of the glutamate transporters, the neutral amino acid transport system ASCT, and the insulin-activable neutral/anionic amino acid transporter. Chromosomal assignment studies with somatic Cell hybrid analysis and fluorescent in situ hybridization have located the ATBo gene to human chromosome 19q13.3.

  • modulation of the activity of amino acid transport system l by phorbol esters and calmodulin antagonists in a human placental Choriocarcinoma Cell Line
    Biochimica et Biophysica Acta, 1992
    Co-Authors: Sammanda Ramamoorthy, Frederick H Leibach, Virendra B. Mahesh, Vadivel Ganapathy
    Abstract:

    We investigated the regulation of the activity of amino acid transport system L in the JAR human placental Choriocarcinoma Cell Line by agents which are known to modulate the activities of three different classes of protein kinases, A-kinase, C-kinase and CaM-kinase. The system L activity was measured by determining the uptake of leucine in these Cells, grown as confluent monolayers. Leucine uptake in these Cells was predominantly Na(+)-independent, was stimulated by lowering the extraCellular pH and was inhibited by hydrophobic neutral amino acids. These characteristics demonstrate that uptake of leucine in this Cell Line occurs primarily via system L. Treatment of the Cells with cholera toxin and forskolin, agents which are known to elevate intraCellular cAMP levels, did not have any effect on the activity of system L. 4 beta-Phorbol 12-myristate 13-acetate, an activator of C-kinase, but not the inactive analogue 4 alpha-phorbol 12,13-didecanoate, caused a significant stimulation of system L. The involvement of C-kinase in the phorbol ester-induced stimulation was supported by the finding that staurosporine, an inhibitor of C-kinase, effectively blocked the stimulation. Calmodulin antagonists, calmidazolium, W-7 and CGS 9343 B stimulated system L activity markedly. The potency of these antagonists was in the following order: calmidazolium greater than CGS 9343 B greater than W-7. This stimulatory effect was specific for system L because systems A and ASC were not stimulated by these agents. The stimulation caused by these agents was primarily due to an increase in the maximal velocity, the apparent Km of the system being only minimally affected. It is concluded that the activity of amino acid transport system L in the JAR human placental Choriocarcinoma Cell Line is stimulated by C-kinase, inhibited by CaM-kinase and unaffected by A-kinase.

Donruedee Sanguansermsri - One of the best experts on this subject based on the ideXlab platform.

  • jeg 3 Cell culture supernatants cause reduced interferon gamma and interleukin 17 production in mixed lymphocyte reactions
    American Journal of Reproductive Immunology, 2007
    Co-Authors: Sutatip Pongcharoen, Pannika R Niumsup, Donruedee Sanguansermsri
    Abstract:

    Problem Immunoregulatory effects of Choriocarcinoma-derived factors on leukocytes have been documented. The present study was designed to investigate the effect of JEG-3 culture supernatants on interferon-γ (IFN-γ), interleukin-17 (IL-17) and IL-1β production in the mixed lymphocyte reactions (MLRs). Method of study A human Choriocarcinoma Cell Line JEG-3 was used to test the effects of its culture supernatants on the proliferation and cytokine production in the MLRs. The Cell proliferation was assessed using the BrdU incorporation and the amounts of cytokines were measured using enzyme-linked immunosorbent assays. Results The JEG-3 culture supernatants caused significantly reduced IFN-γ and IL-17 production in the MLRs. However, the supernatants did not influence MLR production of IL-1β. Conclusion IFN-γ and IL-17 are mainly produced by activated T Cells but IL-1β is primarily produced by monocytes, thus suggesting that immunoregulatory factors of JEG-3 Cells selectively inhibit cytokine production by activated T Cells rather than that of the monocytes.

  • jeg 3 Cell culture supernatants cause reduced interferon γ and interleukin 17 production in mixed lymphocyte reactions
    American Journal of Reproductive Immunology, 2007
    Co-Authors: Sutatip Pongcharoen, Pannika R Niumsup, Donruedee Sanguansermsri
    Abstract:

    Problem Immunoregulatory effects of Choriocarcinoma-derived factors on leukocytes have been documented. The present study was designed to investigate the effect of JEG-3 culture supernatants on interferon-γ (IFN-γ), interleukin-17 (IL-17) and IL-1β production in the mixed lymphocyte reactions (MLRs). Method of study A human Choriocarcinoma Cell Line JEG-3 was used to test the effects of its culture supernatants on the proliferation and cytokine production in the MLRs. The Cell proliferation was assessed using the BrdU incorporation and the amounts of cytokines were measured using enzyme-linked immunosorbent assays. Results The JEG-3 culture supernatants caused significantly reduced IFN-γ and IL-17 production in the MLRs. However, the supernatants did not influence MLR production of IL-1β. Conclusion IFN-γ and IL-17 are mainly produced by activated T Cells but IL-1β is primarily produced by monocytes, thus suggesting that immunoregulatory factors of JEG-3 Cells selectively inhibit cytokine production by activated T Cells rather than that of the monocytes.

Shingo Fujii - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Human Reproduction vol.5 no.2 pp. 168–174, 1999
    2015
    Co-Authors: Takeshi Hirano, Toshihiro Higuchi, Hiroshi Fujiwara, Hiroshi Katsuragawa, Takuya Inoue, Nobuhiko Kataoka, Kyong Rim Park, Masamichi Ueda, Michiyuki Maeda, Shingo Fujii
    Abstract:

    CD9 is involved in invasion of human trophoblast-like Choriocarcinoma Cell Line, BeWo Cell

  • Human migrating extravillous trophoblasts express a Cell surface peptidase, carboxypeptidase-M.
    Molecular Human Reproduction, 2003
    Co-Authors: Yoshihiro Nishioka, Toshihiro Higuchi, Yukiyasu Sato, Shinya Yoshioka, Keiji Tatsumi, Hiroshi Fujiwara, Shingo Fujii
    Abstract:

    We previously reported that a Cell-surface aminopeptidase, dipeptidyl peptidase IV, is expressed on extravillous trophoblasts (EVT) and suggested the involvement of its enzyme activity in EVT migration. In this study, we examined the expression of another Cell-surface peptidase, carboxypeptidase-M (CP-M), at human embryo implantation sites, which catalyses biologically active peptides at extraCellular sites. CP-M was immunohistochemically detected on syncytiotrophoblast, but not on cytotrophoblasts in floating chorionic villi (9-12 weeks of gestation). At villus-anchoring sites, CP-M was weakly detected on some EVT in the distal part of the Cell column. CP-M was clearly expressed on EVT in the trophoblastic shells and in the maternal vessels. In the decidua, almost all interstitial trophoblasts expressed CP-M. Flow cytometry and RT-PCR showed that CP-M expression was induced on the outgrown EVT in primary villous explant culture. The CP-M induction on cultured EVT under 20% O(2) concentration was significantly higher than that under 1% O(2) concentration. In invasion assays, migration of JEG-3 Cells, a CP-M-bearing human Choriocarcinoma Cell Line, was significantly enhanced by an inhibitor of CP-M, DL-mercaptomethyl-3-guanidino-ethyltiopropanoic acid (MGTA). These findings indicate that CP-M is a differentiation-related molecule for human EVT and suggest that CP-M expression on EVT is partially regulated by tissue oxygen concentration.

  • peripheral blood mononuclear Cells in early pregnancy promote invasion of human Choriocarcinoma Cell Line bewo Cells
    Human Reproduction, 2002
    Co-Authors: Haruto Egawa, Toshihiro Higuchi, Keiji Tatsumi, Hiroshi Fujiwara, Takeshi Hirano, Takahiro Nakayama, Takahide Mori, Shingo Fujii
    Abstract:

    BACKGROUND During the establishment of the maternal blood circulation around the implanting human embryo, maternal peripheral blood mononuclear Cells (PBMC) directly contact trophoblasts. To determine the physiological significance of this interaction, the effects of PBMC obtained from pregnant women on the proliferative and invasive properties of a human Choriocarcinoma Cell Line, BeWo Cells, were examined. METHODS AND RESULTS PBMC were obtained from women in early pregnancy and from women in the secretory phase of the menstrual cycle. PBMC from pregnant women significantly increased the number of invading BeWo Cells in an invasion assay without affecting the proliferation of BeWo Cells (P +/- 0.05). No significant changes were observed in the co-cultures with PBMC from non-pregnant women. The addition of conditioned medium, which was prepared by 2 days of incubation with PBMC from pregnant women, also enhanced BeWo Cell invasion in a dose-dependent manner. Moreover, when PBMC obtained from non-pregnant women were incubated with recombinant HCG (0-10 IU/ml) for 2 days, significant augmentation of the effect on BeWo Cell invasion was observed in the conditioned medium from HCG-treated PBMC (P +/- 0.05). CONCLUSION This study indicated that soluble factor(s) secreted from PBMC promote BeWo Cell invasion. It also showed the possible involvement of HCG in the regulation of BeWo Cell invasion by PBMC. These findings suggest crosstalk between maternal PBMC and trophoblasts via soluble factor(s), which may play an important role in early embryo implantation.

  • cd9 is involved in invasion of human trophoblast like Choriocarcinoma Cell Line bewo Cells
    Molecular Human Reproduction, 1999
    Co-Authors: Takeshi Hirano, Toshihiro Higuchi, Hiroshi Fujiwara, Hiroshi Katsuragawa, Takuya Inoue, Nobuhiko Kataoka, Kyong Rim Park, Masamichi Ueda, Michiyuki Maeda, Shingo Fujii
    Abstract:

    The CD9 molecule is expressed on human extravillous trophoblasts, which invade the endometrium during implantation and placentation. To elucidate the role of CD9 in trophoblastic function, we investigated the expression of CD9 protein and mRNA in BeWo Cells, a human trophoblast-like Choriocarcinoma Cell Line, using immunohistochemistry, Western blotting and reverse transcription‐polymerase chain reaction (RT‐PCR). When BeWo Cells were cultured with anti-CD9 monoclonal antibodies (mAb), their invasion through the extraCellular matrices was significantly enhanced in a dose-dependent manner. Cell proliferation and human chorionic gonadotrophin production were unaffected. On the other hand, culture in the presence of mAb against integrins a3, a5 and b1, which partially block the interaction with the extraCellular matrices, inhibited BeWo Cell invasion. Anti-CD9 monoclonal antibody had a stimulatory effect on BeWo Cell invasion in the presence of anti-integrin a3 antibody. In contrast, it had no effect in the presence of mAb against integrins a5 and b1, which were also highly expressed on BeWo Cells. These findings suggest that CD9 has a function connected with the invasive properties of BeWo Cells, which is partially mediated by integrin a5b1. This may relate to the involvement of CD9 in trophoblastic invasion.

Hui Zhu - One of the best experts on this subject based on the ideXlab platform.

  • Article Regulation of Human Trophoblast GLUT3 Glucose Transporter by Mammalian Target of Rapamycin Signaling
    2016
    Co-Authors: Jiao Wang, Ruotong Zhang, Xin Qian, Hui Zhu
    Abstract:

    Abstract: Glucose transporter isoform-3 (GLUT3), one of the primary placental facilitative glucose transporters responsible for basal glucose transport, has a crucial role in glucose transport and fetal growth during early pregnancy. A GLUT3 mutation in mice has been reported to cause loss of early pregnancy or late-gestational fetal growth restriction. However, the underlying mechanisms that regulate the placental GLUT3 transporter in humans are largely unknown. In the present study, we used the JEG-3 human Choriocarcinoma Cell Line, which resembles a first trimester placental model, to study the role of the mammalian target of rapamycin complex 1 (mTORC1) in the regulation of placental GLUT3. We combined rapamycin treatment and small interfering (si) RNA-mediated silencing approaches with mRNA and protein expression/localization studies to investigate the alteration of GLUT3 expression and localization following mTORC1 inhibition in JEG-3 trophoblasts. Inhibition of mTORC1 signaling by silencing raptor decreased GLUT3 mRNA expression (−41%) and protein expression (−50%). Similar effects were obtained in Cells in which mTORC1 was inhibited by rapamycin. Immunofluorescence analysis revealed that GLUT3 expression was markedly reduced in the Cell surface and cytoplasm of JEG-3 Cells in response to mTORC

  • Regulation of Human Trophoblast GLUT3 Glucose Transporter by Mammalian Target of Rapamycin Signaling
    International journal of molecular sciences, 2015
    Co-Authors: Jiao Wang, Ruotong Zhang, Xin Qian, Hui Zhu
    Abstract:

    Glucose transporter isoform-3 (GLUT3), one of the primary placental facilitative glucose transporters responsible for basal glucose transport, has a crucial role in glucose transport and fetal growth during early pregnancy. A GLUT3 mutation in mice has been reported to cause loss of early pregnancy or late-gestational fetal growth restriction. However, the underlying mechanisms that regulate the placental GLUT3 transporter in humans are largely unknown. In the present study, we used the JEG-3 human Choriocarcinoma Cell Line, which resembles a first trimester placental model, to study the role of the mammalian target of rapamycin complex 1 (mTORC1) in the regulation of placental GLUT3. We combined rapamycin treatment and small interfering (si) RNA-mediated silencing approaches with mRNA and protein expression/localization studies to investigate the alteration of GLUT3 expression and localization following mTORC1 inhibition in JEG-3 trophoblasts. Inhibition of mTORC1 signaling by silencing raptor decreased GLUT3 mRNA expression (−41%) and protein expression (−50%). Similar effects were obtained in Cells in which mTORC1 was inhibited by rapamycin. Immunofluorescence analysis revealed that GLUT3 expression was markedly reduced in the Cell surface and cytoplasm of JEG-3 Cells in response to mTORC1 silencing. Because placental mTORC1 activity and GLUT3 expression are decreased in human intrauterine growth restriction, our data suggested one possible mechanism for the abnormal fetal growth in this pregnancy complication.