Differential Expression of SERPINs in the Chicken Oviduct

Whasun Lim(임화선),Jihye Kim,Suzie E. Ahn,Wooyoung Jeong,Jinyoung Kim,Jin Won Choi,Jae Yong Han,Fuller W. Bazer,Gwonhwa Song 한국가금학회 2010 한국가금학회 정기총회 및 학술발표회 Vol.27 No.-

Serpins are a superfamily of relating to inhibit protease with common structural features and inhibitory mechanisms. It is also known as ov-serpins in chicken which are ovalbumin and secreted chicken oviduct protein. However, to our knowledge, it is relatively little known about serpins in chickens. In this study, we investigated the expression patterns of serpin genes in the adult chicken oviduct and also the neonatal oviduct of 37-day-old chicks which were treated with an estrogen analogue, diethylstilbestrol (DES). As results SERPINB3 and SERPINB11 genes were highly expressed in DES-treated oviducts but not in non-treated oviduct. Localization of SERPINB3 and SERPINB11 transcripts were specifically found in the gland-like area of adult chicken oviduct. Collectively, our current data suggest that SERPINB3 and SERPINB11 may have distinct functions in the differentiation processes dependent of steroid hormone regulation in the hen oviduct.

Trichlorfon inhibits proliferation and promotes apoptosis of porcine trophectoderm and uterine luminal epithelial cells

Lim, Whasun,An, Yikyung,Yang, Changwon,Bazer, Fuller W.,Song, Gwonhwa Elsevier 2018 Environmental pollution Vol.242 No.1

<P><B>Abstract</B></P> <P>Trichlorfon is an organophosphate insecticide widely used in agriculture. Additionally, it is applied to pigs for control of endo- and ectoparasites. Previous studies have shown the effects of trichlorfon in pigs during late stages of gestation; however, little is known about its effects during early pregnancy, including implantation and placentation. We investigated whether trichlorfon affects proliferation and apoptosis of porcine trophectoderm (pTr) and uterine luminal epithelial (pLE) cells. Trichlorfon inhibited the proliferation of pTr and pLE cells, as evidenced by cell cycle arrest, and altered the expression of proliferation-related proteins. In addition, trichlorfon induced cell death and apoptotic features, such as loss of mitochondrial membrane potential and DNA fragmentation, in pTr and pLE cells. Moreover, trichlorfon treatment decreased concentrations of Ca<SUP>2+</SUP> in the cytoplasm in both cell lines and increased concentrations of Ca<SUP>2+</SUP> in mitochondria of pTr cells. Trichlorfon inhibited the activation of phosphoinositide 3-kinase/AKT and mitogen-activated protein kinase signaling pathways in pTr and pLE cells. Therefore, we suggest that trichlorfon-treated pTr and pLE cells exhibited abnormal cell physiology which might lead to early pregnancy failure.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Trichlorfon induces apoptosis and cell cycle arrest of porcine trophectoderm cells. </LI> <LI> Trichlorfon-induced oxidative stress results in mitochondrial membrane disruption. </LI> <LI> Trichlorfon inhibits the normal growth and survival of porcine trophoblast cells. </LI> <LI> Trichlorfon might lead to early pregnancy failure in pigs. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>A schematic diagram of mechanisms responsible for proapoptotic and antiproliferative effects of trichlorfon on pTr and pLE cells. Trichlorfon decreased the phosphorylation of PI3K/AKT and MAPK signaling proteins, which are normally activated during cell proliferation. Also, trichlorfon decreased the expression of PCNA and phosphorylation of cyclin D1 resulting in cell cycle arrest. In addition, a mitochondrial Ca<SUP>2+</SUP> overload induced by trichlorfon in pTr cells may be responsible for the decrease in mitochondrial membrane potential (MMP, Δψm). Loss of Δψm subsequently promoted apoptosis in pTr and pLE cells. Overall, exposure to trichlorfon decreased the viability of pTr and pLE cells which may cause pregnancy failure.</P> <P>[DISPLAY OMISSION]</P>

Functional Role of Polyphenols in Reproductive Health and Early Pregnancy

Whasun Lim,Gwonhwa Song 한국식품영양과학회 2016 한국식품영양과학회 학술대회발표집 Vol.2016 No.10

SERPINB3 in the Chicken Model of Epithelial Ovarian Cancer

Whasun Lim,Hee Seung Kim,Jinyoung Kim,Wooyoung Jeong,Jin Young Lee,Chul Hong Lim,Seung Min Bae,Jae Yong Han,Fuller W. Bazer,Gwonhwa Song 한국동물번식학회 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s

SERPINB3 (also known Squamous cell carcinoma antigen 1, SCCA1) is involved in apoptosis, immune response, cell migration and invasiveness of cells. It has been investigated in various types of squamous cell carcinoma. Therefore we investigated the functional role of SERPINB3 gene in human epithelial ovarian cancer (EOC) using laying hens, the most relevant animal model. In 136 laying hens, EOC was found in 10 (7.4%). We compared the expression and localization of SERPINB3 using RT-PCR, quantitative RT-PCR, in situ hybridization and immunohistochemistry, and SERPINB3 activation was detected in chicken and human ovarian cancer cell lines using immunofluorescence microscopy. Thereafter, we examined the prognostic value of SERPINB3 expression in patients with EOC by multivariate linear logistic regression and Cox’ proportional hazard analyses. In present study, SERPINB3 mRNA was induced in cancerous ovaries (p< 0.01), and it was only expressed in the glandular epithelium(GE) of cancerous ovaries of laying hens. SERPINB3 protein was localized predominantly to the nucleus of glandular epithelium in cancerous ovaries of laying hens, and it was abundant in the nucleus of both chicken and human ovarian cancer cell lines. In 109 human patients with EOC, 15 (13.8%), 66 (60.6%) and 28 (25.7%) of those patients showed weak, moderate and strong expression of SERPINB3 protein, respectively. Strong expression of SERPINB3 protein was a prognostic factor for platinum resistance (adjusted OR, 5.94; 95% Confidence Limits, 1.21-29.15). Therefore SERPINB3 may play an important role in ovarian carcinogenesis and be a novel biomarker for predicting platinum resistance and a poor prognosis for survival in patients with EOC. This research was funded by the World Class University (WCU) program (R31-10056), Basic Science Research Program (2010- 0013078) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology and by the Next-Generation BioGreen 21 Program (No.PJ008142), Rural Development Administration, Republic of Korea.

Inhibitory effects of delphinidin on the proliferation of ovarian cancer cells via PI3K/AKT and ERK 1/2 MAPK signal transduction

Lim, Whasun,Song, Gwonhwa D.A. Spandidos 2017 Oncology letters Vol.14 No.1

<P>Delphinidin is a member of the anthocyanidin family and is a natural pigment in red cabbage, berries, sweet potatoes and grapes. It possesses nutraceutical properties against various chronic diseases and types of cancer. However, little is known about its preventative effects on epithelial ovarian cancer, a disease that is associated with a low survival rate, a poor prognosis and a high rate of recurrence. The results of the present study demonstrated that the proliferation of SKOV3 cells decreased in a dose-dependent manner in response to treatment with delphinidin, and the phosphorylation of carcinogenic protein kinases associated with the progression of epithelial ovarian cancer was affected by delphinidin treatment. The levels of phosphorylated protein kinase B (AKT), ribosomal protein S6 kinase β-1 (P70S6K), ribosomal protein S (S6), extracellular signal-regulated kinase (ERK)1/2 and p38 were suppressed by increasing concentrations of delphinidin. Furthermore, the combination of certain pharmacological inhibitors, including phosphoinositide 3-kinase (PI3K; LY294002), ERK1/2 (U0126) and delphinidin significantly reduced the proliferation of SKOV3 cells and the phosphorylation of each of those target proteins. In addition, delphinidin treatment exerted anti-proliferative effects on paclitaxel-resistant SKOV3 cells, compared with treatment with paclitaxel alone. These results indicate that delphinidin inhibits the proliferation of SKOV3 cells through inactivation of PI3K/AKT and ERK1/2 mitogen-activated protein kinase signaling cascades, and that this cell signaling pathway may be a pivotal therapeutic target for the prevention of epithelial ovarian cancer, including paclitaxel-resistant ovarian cancer.</P>

Differential Expression of Gallicin 11 in the Oviduct in Response to Diethylstilbestrol and in Ovarian Carcinomas of Chickens

Whasun Lim,Wooyoung Jeong,Seung-Min Bae,Gahee Jo,Gwonhwa Song 한국동물생명공학회(구 한국동물번식학회) 2013 발생공학 국제심포지엄 및 학술대회 Vol.2013 No.1

Cell-specific expression and signal transduction of C-C motif chemokine ligand 2 and atypical chemokine receptors in the porcine endometrium during early pregnancy

Lim, Whasun,Bae, Hyocheol,Bazer, Fuller W.,Song, Gwonhwa Elsevier 2018 Developmental and comparative immunology Vol.81 No.-

<P><B>Abstract</B></P> <P>Chemokines and atypical chemokine receptors (ACKRs; also known as chemokine decoy receptors) play an important role in reproductive immunology by recruiting leukocytes during early pregnancy. The aim of this study was to determine the expression of C-C motif chemokine ligand 2 (CCL2) and ACKRs in the endometrium during estrous cycle and early pregnancy, and to investigate the functional effects of CCL2 on porcine uterine luminal epithelial (pLE) cells. Our results indicated that CCL2, ACKR1, ACKR3, and ACKR4 were strongly detected in the glandular and luminal epithelium of the endometrium during early pregnancy compared to that in non-pregnant pigs. Recombinant CCL2 improved pLE cell proliferation via activation of the PI3K and MAPK pathways and suppression of endoplasmic reticulum (ER) stress by reducing the expression of ER stress regulatory genes. Collectively, these results provide novel insights into CCL2-mediated signaling mechanisms in the porcine endometrium at the maternal-fetal interface during early pregnancy.</P> <P><B>Highlights</B></P> <P> <UL> <LI> CCL2-ACKR system is regulated in the porcine uterine endometrium. </LI> <LI> CCL2 induces the proliferation and cell cycle progression of porcine uterine cells. </LI> <LI> CCL2 activates PI3K/AKT and MAPK pathways for cellular proliferation in the uterus. </LI> <LI> CCL2 reduces ER stress regulatory gene expression in porcine luminal epithelia cells. </LI> <LI> CCL2 may play an important role in maternal-fetal interaction during early pregnancy. </LI> </UL> </P>

Differential expression of select members of the SLC family of genes and regulation of expression by microRNAs in the chicken oviduct.

Lim, Chul-Hong,Jeong, Wooyoung,Lim, Whasun,Kim, Jinyoung,Song, Gwonhwa,Bazer, Fuller W Society for the Study of Reproduction [etc.] 2012 BIOLOGY OF REPRODUCTION Vol.87 No.6

<P>The yolk and white of eggs from chickens contain proteins and other molecules either secreted or transported by cells of the reproductive tract, or secreted by the liver and transported to the ovarian follicles of laying hens. Nutrients transported by solute carriers (SLCs) include glucose, electrolytes, and amino acids. Although SLC genes have been investigated in mammals, there are few studies of expression of SLC genes in the chicken oviduct. Therefore, we investigated temporal and cell-specific expression of selected SLC genes at 3 h and 20 h postovulation and regulation of their expression by microRNAs (miRs). Expression of SLC1A4 (glutamate and neutral amino acid transporter), SLC13A2 (dicarboxylate transporter), and SLC35B4 (UDP-xylose: UDP-N-acetylglucosamine transporter) mRNAs was limited to glandular epithelium (GE), while SLC4A5 (sodium bicarbonate cotransporter) and SLC7A3 (cationic amino acid transporter) mRNAs were expressed predominantly in the luminal epithelium of the magnum. Interestingly, SLC1A4, SLC4A5, SLC13A2 and SLC35B4 mRNAs were abundant only in GE of the shell gland, whereas SLC7A3 was not detected in the shell gland. In the magnum, SLC7A3 and SLC4A5 were expressed, but SLC1A4, SLC35B4, and SLC13A2 were not expressed at 20 h postovulation. In the shell gland, all SLC mRNAs were expressed at both time points, except for SLC7A3. The miRNA target validation assay revealed that miR-1764 and miR-1700 bind directly to SLC13A2 and SLC35B4 transcripts, respectively, to regulate expression. Results of this study demonstrate cell-specific and temporal changes in expression of selected SLC genes and regulation of SLC13A2 and SLC35B4 expression by miRs in the oviduct of laying hens.</P>

Avian SERPINB11 gene: characteristics, tissue-specific expression, and regulation of expression by estrogen.

Lim, Whasun,Kim, Ji-Hye,Ahn, Suzie E,Jeong, Wooyoung,Kim, Jinyoung,Bazer, Fuller W,Han, Jae Yong,Song, Gwonhwa Society for the Study of Reproduction [etc.] 2011 BIOLOGY OF REPRODUCTION Vol.85 No.6

<P>Serpins, a group of proteins with similar structural and functional properties, were first identified based on their unique mechanism of action: their inhibition of proteases. While most serpins have inhibitory roles, certain serpins are not involved in canonical proteolytic cascades but perform diverse functions including storage of ovalbumin in egg white, transport of hormones (thyroxine- and cortisol-binding globulin), and suppression of tumors. Of these, serpin peptidase inhibitor, clade B, member 11 (SERPINB11) is not an inhibitor of known proteases in humans and mice, and its function is unknown. In the present study, the SERPINB11 gene was cloned, and its expression profile was analyzed in various tissues from chickens. The chicken SERPINB11 gene has an open reading frame of 1346 nucleotides that encode a protein of 388 amino acids that has moderate homology (38.8%-42.3%) to mammalian SERPINB11 proteins. Importantly, SERPINB11 mRNA is most abundant in the chicken oviduct, specifically luminal and glandular epithelia, but it was not detected in any other chicken tissues of either sex. We then determined effects of diethylstilbestrol (DES; a synthetic nonsteroidal estrogen) on SERPINB11 expression in the chicken oviduct. Treatment of young chicks with DES induced SERPINB11 mRNA and protein only in luminal and glandular epithelial cells of the oviduct. Collectively, these results indicate that the novel estrogen-induced SERPINB11 gene is expressed only in epithelial cells of the chicken oviduct and implicate SERPINB11 in regulation of oviduct development and differentiated functions.</P>

Chicken SERPINBs: Characteristics, Tissue Specific Expression and Regulation of Expression by Estrogen

Whasun Lim,Jee-Hye Kim,Suzie E. Ahn,Wooyoung Jeong,Jinyoung Kim,Jae Yong Han,Fuller W. Bazer,Gwonhwa Song 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1

Serpins are a superfamily of related protease inhibitors with common structural features and inhibitory mechanisms. However, SERPINA 14 in mammals does not have inhibitory activity against most known proteases. Rather, it may have an immunoregulatory role in mammals to prevent rejection of the fetal allograft by inhibiting lymphocyte proliferation and natural killer cell function. In the pig, SERPINA14 is involved in iron transport to the fetus by binding to and stabilizing the iron-binding protein uteroferrin (ACP5). In chickens, these very little known about serpins in chickens. Therefore, we investigated the expression patterns of serpin genes in the oviduct of adult hens and in the oviduct of 37-day-old chicks treated with an estrogen analogue, diethylstilbestrol (DES). Results indicated that SERPINB3 and SERPINB11 genes were highly expressed in oviducts of DES-treated chicks, but not in oviducts of control chicks. Both SERPINB3 and SERPINB11 transcripts were localized specifically to the gland-like areas of oviducts of DES-treated chicks. Immunohistochemical analyses confirmed that SERPINB3 and SERPINB11 proteins were present in the gland-like area and luminal epithelium of the oviducts of DES-treated chicks. Collectively, the results suggest that SERPINB3 and SERPINB11 are expressed in response to estrogens and they have distinct functions related to development and differentiation of the mature oviduct in hens.